Uses of Soluble Corn Fiber for Increasing Colonic Bacteria Populations and Increasing Mineral Absorption

ABSTRACT

The present invention relates to fermentable soluble fibers, such as soluble corn fiber (SCF), and its uses in increasing colonic bacteria populations, and edible compositions useful in increasing colonic bacteria populations.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority of U.S. ProvisionalPatent Application Ser. No. 61/804,584, filed Mar. 22, 2013, which ishereby incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates generally to fermentable soluble fibers,such as soluble corn fiber (SCF), and uses and compositions thereof. Incertain aspects, the present invention relates to methods of increasingcolonic bacteria populations in a subject.

Summary of the Related Art

The gut microflorae form a complex ecosystem that interacts with hostcells and nutrients. An adult human body contains a living bacterialbiomass of greater than 10¹⁴ and more than 400 different species, whichrepresents the largest, densest, and most diverse microbial community inthe human body. The presence of the gut bacteria is a part of normalhuman physiology and is important for the development of gut functions,harvesting energy from dietary carbohydrates, harvesting essentialvitamins and metabolizing environmental chemicals in the gut. Recentstudies further suggested that gut bacteria may be involved in fatstorage and affect weight gain and loss. Gut bacteria is also involvedin maturation of the immune system, is in constant communication withthe immune system, and protection against pathogens. Given theimportance of gut bacteria in health and wellness, a strong interest infunctional food ingredients to enhance the populations of beneficial gutbacteria has emerged.

Adolescence is an important life-stage for bone health providing aunique opportunity to maximize mineral retention and prevent the risk ofosteoporosis-related fractures later in life. Because calcium isbecoming increasingly deficient in the diet due to decreasing milkconsumption, a strong interest in functional food ingredients to enhancecalcium utilization has emerged.

SUMMARY OF THE INVENTION

In one broad aspect, the invention provides a method of increasing oneor more colonic bacteria populations in a subject, the method comprisingorally administering to the subject a composition comprising afermentable soluble fiber. In another aspect, the invention provides amethod of increasing one or more colonic bacteria populations in asubject, the method comprising orally administering to the subject acomposition comprising soluble corn fiber.

In one aspect the invention provides a method of increasing one or morecolonic bacteria populations selected from the genera Parabacteroides,Butyricicoccus, Oscillibacter, and Dialister in a subject, the methodcomprising orally administering to the subject a composition comprisinga fermentable soluble fiber. In another aspect the invention provides amethod of increasing one or more colonic bacteria populations selectedfrom Parabacteroides, Butyricicoccus, Oscillibacter, and Dialister in asubject, the method comprising orally administering to the subject acomposition comprising soluble corn fiber.

In another aspect the invention provides a method of increasing one ormore colonic bacteria populations selected from the genera Bacteroides,Butyricicoccus, Oscillibacter, and Dialister in a subject, the methodcomprising orally administering to the subject a composition comprisinga fermentable soluble fiber (e.g., soluble corn fiber).

In another aspect the invention provides a method of increasing one ormore colonic bacteria populations selected from the generaParabacteroides, Bifidobacterium, Alistipes, Anaerococcus,Catenibacterium, genera within the order Clostridiales, and generawithin the family Ruminococcaceae in a subject, the method comprisingorally administering to the subject a composition comprising afermentable soluble fiber (e.g., soluble corn fiber).

In another aspect the invention provides a method of increasing one ormore colonic bacteria populations selected from the generaParabacteroides, Dialister, Akkermansia, and genera within the familyLachnospiraceae in a subject, the method comprising orally administeringto the subject a composition comprising a fermentable soluble fiber(e.g., soluble corn fiber).

In another aspect, the invention provides a method of increasing one ormore colonic bacteria populations in a subject, the method comprisingorally administering to the subject a composition comprising afermentable soluble fiber, such as soluble corn fiber, at least about 3g/day, at least about 5 g/day, at least about 10 g/day, at least about15 g/day, at least about 20 g/day or even at least about 25 g/day.

In another aspect, the invention provides a method of increasing one ormore colonic bacteria populations in a subject, the method comprisingorally administering to the subject a composition comprising afermentable soluble fiber, such as soluble corn fiber such that there isa decrease in fecal pH to a value below about 5.5 (e.g., a decrease infecal pH from about 7 to about 4.5). Such decrease, can for example,result in an increase in the bioavailability of calcium.

In another aspect, the invention provides a method of decreasing fecalpH to a value no more than about 5.5 (e.g., to a fecal pH of about 4.5),the method comprising orally administering to the subject a compositioncomprising a fermentable soluble fiber, such as soluble corn fiber.

In another aspect, the invention provides a method of increasing mineral(e.g., calcium, iron, zinc, copper, potassium and/or magnesium)absorption in a subject, the method comprising orally administering tothe subject a composition comprising a fermentable soluble fiber, suchas soluble corn fiber. In certain embodiments of the methods andcompositions as described herein, the mineral is absorbed as a divalentcation. In certain embodiments of the methods and compositions asdescribed herein, the mineral is calcium. In other embodiments of themethods and compositions as described herein, the mineral is calciumand/or magnesium. In certain embodiments of the methods and compositionsas described herein, the mineral is calcium and/or iron. In otherembodiments of the methods and compositions as described herein, themineral is calcium, magnesium and/or iron.

In another aspect, the invention provides a method of increasing mineral(e.g., calcium, iron, zinc, copper, potassium/or and magnesium, asdescribed above) absorption in a subject, the method comprising orallyadministering to the subject a composition comprising a fermentablesoluble fiber, such as soluble corn fiber, at a rate of at least about 3g/day, at least about 5 g/day, at least about 10 g/day, at least about12 g/day, at least about 15 g/day, at least about 20 g/day or even atleast about 25 g/day.

In another aspect, the invention provides an edible product comprising afermentable soluble fiber, such as soluble corn fiber, and one or morebacterial populations selected from the group consisting ofLactobacillus, Bacteroides, Parabacteroides, Alistipes, Bifidobacterium,Butyricicoccus, Oscillibacter, Dialister and any combinations thereof.

In another aspect the invention provides an edible compositioncomprising one or more (e.g., two or more or three or more) bacteriapopulations selected from the genera (e.g., each selected from adifferent genus) Bacteroides, Butyricicoccus, Oscillibacter, andDialister. The edible composition can optionally include a fermentablesoluble fiber (e.g., soluble corn fiber).

In another aspect the invention provides an edible compositioncomprising one or more (e.g., two or more or three or more) bacteriapopulations selected from the genera (e.g., each selected from adifferent genus) Parabacteroides, Dialister, Akkermansia, and generawithin the family Lachnospiraceae. The edible composition can optionallyinclude a fermentable soluble fiber (e.g., soluble corn fiber).

In another aspect the invention provides an edible compositioncomprising one or more (e.g., two or more or three or more) bacteriapopulations selected from the genera (e.g., each selected from adifferent genus) Parabacteroides, Bifidobacterium, Alistipes,Anaerococcus, Catenibacterium, genera within the order Clostridiales;and genera within the family Ruminococcaceae. The edible composition canoptionally include a fermentable soluble fiber (e.g., soluble cornfiber).

BRIEF DESCRIPTION OF THE DRAWINGS

The invention set forth herein can be advantageously understood withregard to the drawings.

FIG. 1 shows the effect of SCF on fractional calcium absorption(mean+SEM) during Day 1 and Day 2 following a calcium absorption testwith dual stable isotopes in early adolescent boys and girls. A generallinear model that included treatment, sequence, and phase for each timeperiod (0-24 h and 24-48 h) indicated that calcium absorption for SCFwas higher than that for control at 24-48 h (*P=0.02) but not at 0-24 h(P=0.09).

FIG. 2 shows comparison of SCF and Control treatments on fractionalcalcium absorption measured by 0-24 h and 24-48 h urine collections.

FIG. 3 shows a comparison of average relative proportions of bacterialfamilies in subjects at the beginning (B) and end (E) of clinicalsessions where diets included soluble corn fiber (SCF) vs control (Con).Only families representing >1.0% of the total community in at least onetreatment are depicted. Error bars represent standard errors of means.Letters depict significant differences within each family (p<0.05).

FIG. 4 shows a histogram comparing average proportion of major bacterialphyla at the beginning (B) and end (E) of each SCF diet treatment.

FIG. 5 shows the rarefaction analysis of Chao1 diversity measures madefrom beginning (B) and end (E) fecal samples collected from subjects ondifferent SCF diet treatment.

FIG. 6 shows Principal Coordinate Analysis (PCoA) of Jackknife BrayCurtis distances (normalized Manhattan distance) of communitycomposition coded SCF diet supplement samples collected at the beginning(B) and end (E) of the SCF treatment.

FIG. 7 shows Principal Coordinate Analysis (PCoA) of Jackknife BrayEuclidean distances of community composition coded SCF diet supplementsamples collected at the beginning (B) and end (E) of the SCF treatment.

FIG. 8 shows Principal Coordinate Analysis (PCoA) of Jackknife Analysisof Unifrac G phylogenetic distances of community composition collectedfrom subjects at the beginning (B) and end (E) of the SCF treatment.

FIG. 9 is a schematic diagram demonstrating an example of a method formaking fermentable soluble fiber.

DETAILED DESCRIPTION

Before the disclosed methods and materials are described, it is to beunderstood that the aspects described herein are not limited to specificembodiments, methods, or compositions, and as such can, of course, vary.It is also to be understood that the terminology used herein is for thepurpose of describing particular aspects only and, unless specificallydefined herein, is not intended to be limiting.

Throughout this specification, unless the context requires otherwise,the word “comprise” and “include” and variations (e.g., “comprises,”“comprising,” “includes,” “including”) will be understood to imply theinclusion of a stated component, feature, element, or step or group ofcomponents, features, elements or steps but not the exclusion of anyother integer or step or group of integers or steps.

As used in the specification and the appended claims, the singular forms“a,” “an” and “the” include plural referents unless the context clearlydictates otherwise.

Ranges can be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, another aspect includes from the one particular value and/orto the other particular value. Similarly, when values are expressed asapproximations, by use of the antecedent “about,” it will be understoodthat the particular value forms another aspect. It will be furtherunderstood that the endpoints of each of the ranges are significant bothin relation to the other endpoint, and independently of the otherendpoint.

In view of the present disclosure, the methods and compositionsdescribed herein can be configured by the person of ordinary skill inthe art to meet the desired need. In general, the disclosed methods andcompositions provide improvements in gut microbiota. For example, incertain aspects, the methods of the disclosure increase one or morecolonic bacteria populations that are capable of fermentation and shortchain fatty acid production.

For example, in certain embodiments of the methods and compositionsdescribed herein, administering a composition comprising a fermentablesoluble fiber, such as soluble corn fiber increases the population ofone or more colonic bacteria populations, each from a genus selectedfrom the group consisting of Parabacteroides, Butyricicoccus,Oscillibacter, and Dialister, and any combination thereof. For example,in one embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Parabacteroides.In another embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Butyricicoccus. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Oscillibacter. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Dialister. Forexample, in certain embodiments of the methods and compositionsdescribed herein, administering a composition comprising a fermentablesoluble fiber, e.g., soluble corn fiber, increases the population ofParabacteroides and Butyricicoccus; Parabacteroides and Oscillibacter;Parabacteroides and Dialister; Butyricicoccus and Oscillibacter;Butyricicoccus and Dialister, Oscillibacter and Dialister;Parabacteroides, Butyricicoccus and Oscillibacter, Parabacteroides,Butyricicoccus and Dialister; Parabacteroides, Oscillibacter, andDialister, Butyricicoccus, Oscillibacter, and Dialister; orParabacteroides, Butyricicoccus, Oscillibacter, and Dialister. Ofcourse, other bacterial populations can additionally be increased. Incertain such embodiments, absorption of calcium also increases (e.g., asdescribed below).

For example, in certain embodiments of the methods and compositionsdescribed herein, administering a composition comprising a fermentablesoluble fiber, such as soluble corn fiber increases the population ofone or more colonic bacteria populations, each from a genus selectedfrom the group consisting of Bacteroides, Butyricicoccus, Oscillibacter,and Dialister, and any combination thereof. For example, in oneembodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Bacteroides. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Butyricicoccus. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Oscillibacter. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Dialister. Forexample, in certain embodiments of the methods and compositionsdescribed herein, administering a composition comprising a fermentablesoluble fiber, e.g., soluble corn fiber, increases the population ofBacteroides and Butyricicoccus; Bacteroides and Oscillibacter,Bacteroides and Dialister; Butyricicoccus and Oscillibacter;Butyricicoccus and Dialister; Oscillibacter and Dialister, Bacteroides,Butyricicoccus and Oscillibacter, Bacteroides, Butyricicoccus andDialister, Bacteroides, Oscillibacter, and Dialister; Butyricicoccus,Oscillibacter, and Dialister, or Bacteroides, Butyricicoccus,Oscillibacter, and Dialister. Of course, other bacterial populations canadditionally be increased. In certain such embodiments, absorption ofcalcium also increases (e.g., as described below).

In other embodiments of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of one or more colonicbacteria populations, each from a genus selected from the groupconsisting of Parabacteroides, Bifidobacterium, Alistipes, Anaerococcus,Catenibacterium, genera within the order Clostridiales (e.g., notClostridium, Anaerofustis, Anaerococcus, Coprococcus,Peptostreptococcaceae, Sporacetigenium); and genera within the familyRuminococcaceae and any combination thereof. For example, in oneembodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Parabacteroides.In another embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of Bifidobacterium. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of Alistipes. In anotherembodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of Anaerococcus. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of Catenibacterium. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of the familyRuminococcaceae. In another embodiment of the methods and compositionsdescribed herein, administering a composition comprising a fermentablesoluble fiber, such as soluble corn fiber increases the population ofthe order Clostridiales. For example, in certain embodiments of themethods and compositions described herein, administering a compositioncomprising a fermentable soluble fiber, e.g., soluble corn fiber,increases the population of Parabacteroides and Bifidobacterium;Parabacteroides and Alistipes; Parabacteroides and Anaerococcus;Parabacteroides and Catenibacterium; Parabacteroides andRuminococcaceae; Parabacteroides and Clostridiales; Bifidobacterium andAlistipes; Bifidobacterium and Anaerococcus; Bifidobacterium andCatenibacterium; Bifidobacterium and Ruminococcaceae; Bifidobacteriumand Clostridiales; Alistipes and Anaerococcus; Alistipes andCatenibacterium; Alistipes and Ruminococcaceae; Anaerococcus andCatenibacterium; Anaerococcus and Ruminococcaceae; Anaerococcus andClostridiales; Catenibacterium and Ruminococcaceae; Catenibacterium andClostridiales; Ruminococcaceae and Clostridiales; Parabacteroides,Bifidobacterium and Alistipes; Parabacteroides, Bifidobacterium andAnaerococcus; Parabacteroides, Bifidobacterium and Catenibacterium;Parabacteroides, Bifidobacterium and Clostridiales; Parabacteroides,Bifidobacterium and Ruminococcaceae; Parabacteroides, Alistipes andAnaerococcus; Parabacteroides, Alistipes and Catenibacterium;Parabacteroides, Alistipes and Clostridiales; Parabacteroides, Alistipesand Ruminococcaceae; Parabacteroides, Anaerococcus and Catenibacterium;Parabacteroides, Anaerococcus and Clostridiales; Parabacteroides,Anaerococcus and Ruminococcaceae; Parabacteroides, Catenibacterium andClostridiales; Parabacteroides, Catenibacterium and Ruminococcaceae;Parabacteroides, Clostridiales and Ruminococcaceae; Bifidobacterium,Alistipes and Anaerococcus; Bifidobacterium, Alistipes andCatenibacterium; Bifidobacterium, Alistipes and Clostridiales;Bifidobacterium, Alistipes and Ruminococcaceae; Bifidobacterium,Anaerococcus and Catenibacterium; Bifidobacterium, Anaerococcus andClostridiales; Bifidobacterium, Anaerococcus and Ruminococcaceae;Bifidobacterium, Catenibacterium and Clostridiales; Bifidobacterium,Catenibacterium and Ruminococcaceae; Bifidobacterium, Clostridiales andRuminococcaceae; Alistipes, Anaerococcus and Catenibacterium; Alistipes,Anaerococcus and Clostridiales; Alistipes, Anaerococcus andRuminococcaceae; Alistipes, Catenibacterium and Clostridiales;Alistipes, Catenibacterium and Ruminococcaceae; Alistipes, Clostridialesand Ruminococcaceae; Anaerococcus, Catenibacterium and Clostridiales;Anaerococcus, Catenibacterium and Ruminococcaceae; Anaerococcus,Clostridiales and Ruminococcaceae; Catenibacterium, Clostridiales andRuminococcaceae; Parabacteroides, Bifidobacterium, Alistipes andAnaerococcus; Parabacteroides, Bifidobacterium, Alistipes andCatenibacterium; Parabacteroides, Bifidobacterium, Alistipes andClostridiales; Parabacteroides, Bifidobacterium, Alistipes andRuminococcaceae; Parabacteroides, Bifidobacterium, Anaerococcus andCatenibacterium; Parabacteroides, Bifidobacterium, Anaerococcus andClostridiales; Parabacteroides, Bifidobacterium, Anaerococcus andRuminococcaceae; Parabacteroides, Bifidobacterium, Catenibacterium andClostridiales; Parabacteroides, Bifidobacterium, Catenibacterium andRuminococcaceae; Parabacteroides, Bifidobacterium, Clostridiales andRuminococcaceae; Parabacteroides, Alistipes, Anaerococcus andCatenibacterium; Parabacteroides, Alistipes, Anaerococcus andClostridiales; Parabacteroides, Alistipes, Anaerococcus andRuminococcaceae; Parabacteroides, Alistipes, Catenibacterium andClostridiales; Parabacteroides, Alistipes, Catenibacterium andRuminococcaceae; Parabacteroides, Alistipes, Clostridiales andRuminococcaceae; Parabacteroides, Anaerococcus, Catenibacterium andClostridiales; Parabacteroides, Anaerococcus, Catenibacterium andRuminococcaceae; Parabacteroides, Anaerococcus, Clostridiales andRuminococcaceae; Parabacteroides, Catenibacterium, Clostridiales andRuminococcaceae; Bifidobacterium, Alistipes, Anaerococcus andCatenibacterium; Bifidobacterium, Alistipes, Anaerococcus andClostridiales; Bifidobacterium, Alistipes, Anaerococcus andRuminococcaceae; Bifidobacterium, Alistipes, Catenibacterium andClostridiales; Bifidobacterium, Alistipes, Catenibacterium andRuminococcaceae; Bifidobacterium, Alistipes, Clostridiales andRuminococcaceae; Bifidobacterium, Anaerococcus, Catenibacterium andClostridiales; Bifidobacterium, Anaerococcus, Catenibacterium andRuminococcaceae; Bifidobacterium, Anaerococcus, Clostridiales andRuminococcaceae; Bifidobacterium, Catenibacterium, Clostridiales andRuminococcaceae; Alistipes, Anaerococcus, Catenibacterium andClostridiales; Alistipes, Anaerococcus, Catenibacterium andRuminococcaceae; Alistipes, Anaerococcus, Clostridiales andRuminococcaceae; Alistipes, Catenibacterium, Clostridiales andRuminococcaceae; or Anaerococcus, Catenibacterium, Clostridiales andRuminococcaceae. Of course, the person of ordinary skill in the art willappreciate that any combination of 5, 6, or 7 colonic bacteriapopulations, each from a different genus selected from the groupconsisting of Parabacteroides, Bifidobacterium, Alistipes, Anaerococcus,Catenibacterium, genera within the order Clostridiales; and generawithin the family Ruminococcaceae, may be increased by the methodsdescribed herein. Of course, other bacterial populations canadditionally be increased.

In other embodiments of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of one or more colonicbacteria populations, each from a genus selected from the groupconsisting of Parabacteroides, Dialister, Akkermansia, and genera withinthe family Lachnospiraceae (e.g., not Lachnospira). For example, in oneembodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber,e.g., soluble corn fiber, increases the population of Parabacteroides.In another embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of Dialister. In anotherembodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of Akkermansia. Inanother embodiment of the methods and compositions described herein,administering a composition comprising a fermentable soluble fiber, suchas soluble corn fiber increases the population of Lachnospiraceae. Forexample, in certain embodiments of the methods and compositionsdescribed herein, administering a composition comprising a fermentablesoluble fiber, e.g., soluble corn fiber, increases the population ofParabacteroides and Dialister, Parabacteroides and Akkermansia;Parabacteroides and Lachnospiraceae; Dialister and Akkermansia;Dialister and Lachnospiraceae; Akkermansia and Lachnospiraceae;Parabacteroides, Dialister, and Akkermansia; Parabacteroides, Dialister,and Lachnospiraceae; Parabacteroides, Akkermansia, and Lachnospiraceae;Dialister, Akkermansia, and Lachnospiraceae; or Parabacteroides,Dialister, Akkermansia, and Lachnospiraceae. Of course, other bacterialpopulations can additionally be increased.

In certain embodiments of the methods and compositions described herein,one or more of the colonic bacteria populations (e.g., as describedabove) are increased by at least about 5%, at least about 10%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 50%, at least about 60%, at leastabout 80%, or even at least about 100% as compared to a non-treatedsubject. In certain such embodiments, the colonic bacteria population isincreased by no more than about 500%. In other such embodiments, thecolonic bacteria population is increased by no more than about 400%. Inother such embodiments, the colonic bacteria population is increased byno more than about 300%. In other such embodiments, the colonic bacteriapopulation is increased by no more than about 200%. In other suchembodiments, the colonic bacteria population is increased by no morethan about 100%. In certain embodiments of the methods and compositionsdescribed herein, each of the one or more of the colonic bacteriapopulations (e.g., as described above) are increased by at least about5%, at least about 10%, at least about 20%, at least about 50%, or evenat least about 100% as compared to a non-treated subject. This meansthat there are instances where each of these bacteria may be affectedindependently of each other at different rates (e.g., one bacteria mayincrease by 50% in population, whereas another bacteria may onlyincrease 25%). In certain such embodiments, each colonic bacteriapopulation is increased by no more than about 500%. In other suchembodiments, each colonic bacteria population is increased by no morethan about 400%. In other such embodiments, each colonic bacteriapopulation is increased by no more than about 300%. In other suchembodiments, each colonic bacteria population is increased by no morethan about 200%. In other such embodiments, each colonic bacteriapopulation is increased by no more than about 100%.

In certain embodiments of the methods and compositions described herein,the proportion of one or more of the colonic bacteria populations (e.g.,as described above) as a percentage of total colonic bacteria isincreased by at least about 20%, at least about 25%, at least about 50%,at least about 100%, at least about 200% or even at least about 300% ascompared to a non-treated subject. In certain such embodiments, theproportion of one or more of the colonic bacteria populations as apercentage of total colonic bacteria is increased by no more than about700%. In other such embodiments, the proportion of one or more of thecolonic bacteria populations as a percentage of total colonic bacteriais increased by no more than about 600%. In other such embodiments, theproportion of one or more of the colonic bacteria populations as apercentage of total colonic bacteria is increased by no more than about500%. In other such embodiments, the proportion of one or more of thecolonic bacteria populations as a percentage of total colonic bacteriais increased by no more than about 400%. In certain embodiments of themethods and compositions described herein, the proportion (i.e., as apercentage of total colonic bacteria) of each of the one or more of thecolonic bacteria populations (e.g., as described above) is increased byat least about 20%, at least about 25%, at least about 50%, at leastabout 100%, at least about 200% or even at least about 300% as comparedto a non-treated subject. This means that there are instances where eachof these bacteria may be affected independently of each other atdifferent rates (e.g., one bacteria population may increase by 50% inproportion, whereas another bacteria population may only increase 25%).In certain such embodiments, each proportion is increased by no morethan about 500%. In other such embodiments, each proportion is increasedby no more than about 400%. In other such embodiments, each proportionis increased by no more than about 300%. In other such embodiments, eachproportion is increased by no more than about 200%. In other suchembodiments, each proportion is increased by no more than about 100%.

In another embodiment, a method of increasing one or more colonicbacteria populations in a subject includes orally administering to thesubject a fermentable soluble fiber, such as soluble corn fiber. Incertain such embodiments, the oral administration is performed such thatthere is a decrease in fecal pH (e.g., as described below, to a value nomore than about 5.5, for example, from a pH value of about 7 to a pHvalue of about 4.5). Such decrease, can for example, result in anincrease in the bioavailability of minerals (e.g., divalent mineralssuch as calcium, as described above).

In another embodiment, the methods of the disclosure also decrease fecalpH in a subject by orally administering to the subject a fermentablesoluble fiber, such as soluble corn fiber. For example, in certainembodiments of the methods and compositions as described herein, fecalpH is reduced by at least about 1.5 pH units, at least about 2 pH units,or even by at least about 2.5 pH units as compared to a non-treatedsubject. In certain embodiments of the methods and compositions asdescribed herein, fecal pH is reduced to no more than about 5.5, no morethan about 5, or even no more than about 4.5. In certain embodiments ofthe methods and compositions as described herein, fecal pH is reduced toa value in the range of about 4 to about 5.5, about 4.5 to about 5.5,about 4 to about 5, or about 4.5 to about 5. In certain embodiments,fecal pH is reduced to about 4.5, for example, from about 7 to about4.5.

In certain embodiments of the methods and compositions as describedhere, the fermentable soluble fiber is soluble corn fiber. Soluble cornfiber is a starch-derived soluble fiber that is made from corn and thatcomprises oligosaccharides that are digestion-resistant,oligosaccharides that are slowly digestible, or a combination thereof.Soluble corn fiber can be made via corn starch hydrolysis, and containsgreater than about 70% fiber and less than about 20% mono- anddisaccharide sugars. The glucose units of the oligosaccharides arelinked primarily by α-1,4 glycosidic bonds, but can also include α-1,6,α-1,3, and α-1,2 bonds.

In certain embodiments of the methods and compositions described herein,the soluble corn fiber has a fiber content in the range of about 70% toabout 100% (w/w). In another embodiment, the fiber content of thesoluble corn fiber is in the range of about 70% to about 90%, or about70% to about 95%, or about 70% to about 100%, about 75% to about 85%, orabout 75% to about 90%, or about 75% to about 95%, or about 75% to about100%, or about 70% to about 85% (w/w). In one embodiment, the fibercontent is about 70% (w/w). In another embodiment, the fiber content isabout 85% (w/w). One of skill in the art will appreciate that fibercontent may be measure by any suitable method known in the art, such asenzymatic gravimetry, liquid chromatography, gas-liquid chromatography,High Pressure Liquid chromatography (HPLC), High Performance AnionExchange Chromatography with Pulsed Amperometric Detection (HPAE-PAD),and other enzymatic and chemical methods. In a preferred embodiment, thefiber content is measured by HPAE-PAD. For example, a Dionex ionchromatograph, DX500, equipped with electrochemical detector andgradient pump, is used to analyze samples that are separated on DionexCarbopac PA1 analytical and guard columns with gradient delivery ofsolvents, detected using a gold electrode with a four-potentialwaveform, and diluted with water and passed through Amicon Ultra-4centrifugal filter devices before analysis.

In certain embodiments of the methods and compositions described herein,the mono- and disaccharide content of the soluble corn fiber is lessthan about 20%. For example, in certain embodiments, the mono- anddisaccharide content of the soluble corn fiber is less than about 15%,less than about 10%, less than about 5%, or even less than about 2%. Incertain such embodiments, the mono- and disaccharide content of thesoluble corn fiber is no less than about 0%, no less than about 0.001%,no less than about 0.01%, or even no less than 0.1%.

In certain embodiments of the methods and compositions described herein,the oligosaccharides of the soluble corn fiber have an average degree ofpolymerization of at least about 5, at least about 7, or at least about9. For example, in certain embodiments of the methods and compositionsdescribed herein, the oligosaccharides of the soluble corn fiber have anaverage degree of polymerization in the range of about 5 to about 20,about 7 to about 20, or about 9 to about 20. In other embodiments, theoligosaccharides of the soluble corn fiber have an average degree ofpolymerization in the range of about 5 to about 15, about 7 to about 15,or about 9 to about 15. For example, in one embodiment the methods andcompositions described herein, the oligosaccharides of the soluble cornfiber have an average degree of polymerization is about 10.

In certain embodiments of the methods and compositions described herein,the oligosaccharide portion of the soluble corn fiber remainssubstantially undigested in the stomach and small intestine of a subjectwhen ingested.

Suitable commercial soluble corn fiber products include PROMITOR™Soluble Corn Fiber 70 (minimum fiber content of about 70%, maximum mono-and disaccharide content of about 20%), and PROMITOR™ Soluble Corn Fiber85 (minimum fiber content of about 85%, maximum mono- and disaccharidecontent of about 2%), available from Tate & Lyle Health & NutritionSciences, Hoffman Estates, Ill.

Certain soluble corn fibers suitable for use in the methods andcompositions described herein are described further in U.S. PatentApplications Publications nos. 2008/0292766, 2006/0210696 and2008/0175977, each of which is hereby incorporated herein by referencein its entirety, and which is attached in the appendix to thisspecification. In certain embodiments of the methods and compositionsdescribed herein, the soluble corn fiber is as described in an aspect orembodiment of U.S. Patent Application Publication no. 2008/0292766,2006/0210696 or 2008/0175977.

Of course, as the person of ordinary skill in the art will appreciate,other fermentable soluble fibers can be used in practicing the methodsand compositions as described herein. In other particular embodiments asdescribed herein, the fermentable soluble fibers is selected frompolydextrose, soluble fiber dextrin (i.e., corn, tapioca, potatostarch), arabinoxylan, arabinoxylan oligosaccharides, xylose, slowlydigestible (digestion resistant) carbohydrates and oligosaccharides, andfunctional combinations thereof, optionally in combination with solublecorn fiber. While certain embodiments of the invention are describedherein with reference to soluble corn fibers, the person of ordinaryskill in the art will appreciate that other fermentable soluble fiberscould be used in place of the soluble corn fibers in certain embodimentsof the invention.

In certain embodiments of the methods and compositions as describedherein, the fermentable soluble fiber, e.g., the soluble corn fiber, isproduced by a process described in U.S. Pat. Nos. 7,608,436, and8,057,840, each of which is hereby incorporated herein by reference inits entirety. For example, in one embodiment, the process to produce thefermentable soluble fiber includes uses an aqueous feed composition thatcomprises at least one monosaccharide or linear saccharide oligomer, andthat has a solids concentration of at least about 70% by weight. Thefeed composition is heated to a temperature of at least about 40° C.,and is contacted with at least one catalyst that accelerates the rate ofcleavage or formation of glucosyl bonds for a time sufficient to causeformation of non-linear saccharide oligomers. In one particularembodiment, the process includes heating an aqueous feed compositionthat comprises at least one monosaccharide or linear saccharideoligomer, and that has a solids concentration of at least about 70% byweight, to a temperature of at least about 40° C.; and contacting thefeed composition with at least one catalyst that accelerates the rate ofcleavage or formation of glucosyl bonds for a time sufficient to causeformation of non-linear saccharide oligomers, wherein a productcomposition is produced that contains a higher concentration ofnon-linear saccharide oligomers than linear saccharide oligomers;wherein the product composition comprises non-linear saccharideoligomers having a degree of polymerization of at least three in aconcentration of at least about 20% by weight on a dry solids basis. Incertain such embodiments, the product composition is produced thatcontains a higher concentration of non-linear saccharide oligomers thanlinear saccharide oligomers. In one embodiment of the process, the atleast one catalyst is an enzyme that accelerates the rate of cleavage orformation of glucosyl bonds. In another embodiment of the process, theat least one catalyst is an acid. In some embodiments of the process,acid and enzyme can be used in sequence, with the feed composition firstbeing treated with enzyme and subsequently with acid, or vice versa.

In certain embodiments of the processes as described with respect toU.S. Pat. Nos. 7,608,436, and 8,057,840, the aqueous feed compositionincludes at least one monosaccharide and at least one linear saccharideoligomer, and may contain several of each. In many cases,monosaccharides and oligosaccharides will make up at least about 70% byweight on a dry solids basis of the feed composition. It is generallyhelpful for the starting material to have as high a concentration ofmonosaccharides as possible, in order to maximize the yield of thedesired oligomers. A high solids concentration tends to drive theequilibrium from hydrolysis toward condensation (reversion), therebyproducing higher molecular weight products. Therefore the water contentof the starting material is preferably relatively low. For example, incertain embodiments, the feed composition comprises at least about 75%dry solids by weight. (“Dry solids” is sometimes abbreviated herein as“ds.”) In some cases, the feed composition comprises about 75-90% solidsby weight, which will generally give the appearance of a viscous syrupor damp powder at room temperature.

Examples of suitable starting materials for the processes as describedwith respect to U.S. Pat. Nos. 7,608,436, and 8,057,840 include, but arenot limited to, syrups made by hydrolysis of starch, such as dextrosegreens syrup (i.e., recycle stream of mother liquor from dextrosemonohydrate crystallization), other dextrose syrups, corn syrup, andsolutions of maltodextrin. If the feed composition comprisesmaltodextrin, the process optionally can also include the steps ofhydrolyzing the maltodextrin to form a hydrolyzed saccharide solutionand concentrating the hydrolyzed saccharide solution to at least about70% dry solids to form the feed composition. The concentrating and thecontacting of the feed with the catalyst can occur simultaneously, orthe concentrating can occur prior to contacting the feed compositionwith the catalyst.

In certain embodiments of the processes as described with respect toU.S. Pat. Nos. 7,608,436, and 8,057,840, the feed composition iscontacted with the at least one catalyst for a period of time that canvary. In some cases, the contacting period will be at least about fivehours. In some embodiments of the invention, the feed composition iscontacted with the at least one catalyst for about 15-100 hours. Inother embodiments, shorter contacting times can be used with highertemperatures, in some cases even less than one hour.

In certain embodiments of the processes as described with respect toU.S. Pat. Nos. 7,608,436, and 8,057,840, enzymatic reversion is used toproduce nonlinear oligosaccharides. The enzyme can be, for example, onethat accelerates the rate of cleavage of alpha 1-2, 1-3, 1-4, or 1-6glucosyl bonds to form dextrose residues. One suitable example is aglucoamylase enzyme composition, such as a commercial enzyme compositionthat is denominated as a glucoamylase. It should be understood that sucha composition can contain some quantity of enzymes other than pureglucoamylase, and it should not be assumed that it is in factglucoamylase itself that catalyzes the desired production of nonlinearoligosaccharides. Therefore, the feed composition can be contacted withglucoamylase or any other enzyme that acts on dextrose polymers. Theamount of enzyme can suitably be about 0.5-2.5% by volume of the feedcomposition. In some embodiments of the process, the feed composition ismaintained at about 55-75° C. during the contacting with the enzyme, orin some cases about 60-65° C. At this temperature, depending on thewater content, the material will become a liquid, or a mixture of liquidand solid. Optionally, the reaction mixture can be mixed or agitated todistribute the enzyme. The reaction mixture is maintained at the desiredtemperature for the time necessary to achieve the desired degree ofreversion to non-linear oligomers. In some embodiments of the process,the feed composition is contacted with the enzyme for about 20-100 hoursprior to inactivation of the enzyme, or in some cases, for about 50-100hours prior to inactivation. Techniques for inactivating glucoamylaseare well known in the field. Alternatively, instead of inactivating theenzyme, it can be separated by membrane filtration and recycled.

In certain embodiments of the processes as described with respect toU.S. Pat. Nos. 7,608,436, and 8,057,840, the resulting composition has ahigh concentration of non-linear oligosaccharides, such as isomaltose.This product composition contains a higher concentration of non-linearsaccharide oligomers than linear saccharide oligomers. In some cases,the concentration of non-linear saccharide oligomers in the finalcomposition is at least twice as high as the concentration of linearsaccharide oligomers.

Another embodiment processes as described with respect to U.S. Pat. Nos.7,608,436, and 8,057,840 involves acid reversion of monosaccharides. Thestarting material is the same as described above with respect to theenzyme version of the process. A variety of acids can be used, such ashydrochloric acid, sulfuric acid, phosphoric acid, or a combinationthereof. In some embodiments of the process, acid is added to the feedcomposition in an amount sufficient to make the pH of the feedcomposition no greater than about 4, or in some cases, in an amountsufficient to make the pH of the feed composition about 1.0-2.5, orabout 1.5-2.0. In some embodiments, the solids concentration of the feedcomposition is about 70-90%, the amount of acid added to the feed isabout 0.05%-0.25% (w/w) acid solids on syrup dry solids, and the feedcomposition is maintained at a temperature of about 70-90° C. during thecontacting with the acid. As in the enzyme version of the process, thereaction conditions are maintained for a time sufficient to produce thedesired oligomers, which in some embodiments of the process will beabout 4-24 hours.

In one particular embodiment of the processes described with respect toU.S. Pat. Nos. 7,608,436, and 8,057,840, the solids concentration of thefeed composition is at least about 80% by weight, acid is added to thefeed composition in an amount sufficient to make the pH of thecomposition about 1.8, and the feed composition is maintained at atemperature of at least about 80° C. for about 4-24 hours after it iscontacted with the acid.

In another particular embodiment of the processes described with respectto U.S. Pat. Nos. 7,608,436, and 8,057,840, the solids concentration ofthe feed composition is about 90-100% by weight, and the feedcomposition is maintained at a temperature of at least about 149° C.(300° F.) for about 0.1-15 minutes after it is contacted with the acid.The acid used to treat the feed can be a combination of phosphoric acidand hydrochloric acid (at the same concentrations discussed above). Inone particular embodiment, the contacting of the feed composition withthe acid takes place in a continuous pipe/flow through reactor.

By far the most plentiful glycosidic linkage in starch is the alpha-1,4linkage, and this is the linkage most commonly broken during acidhydrolysis of starch. But acid-catalyzed reversion (condensation) cantake place between any two hydroxyl groups, and, given the large varietyof combinations and geometries available, the probability of analpha-1,4 linkage being formed is relatively small. The human digestivesystem contains alpha amylases which readily digest the alpha-1,4linkages of starch and corn syrups. Replacing these linkages withlinkages unrecognized by enzymes in the digestive system will allow theproduct to pass through to the small intestines largely unchanged. Thesaccharide distributions resulting from acid treatment are believed tobe somewhat different than from enzyme treatment. It is believed thatthese acid-catalyzed condensation products will be less recognizable bythe enzymes in the human gut than enzyme-produced products, andtherefore less digestible.

The acid treatment progresses differently than enzyme treatment. Enzymesrapidly hydrolyze linear oligomers and slowly form non-linear oligomers,whereas with acid the reduction in linear oligomers and the increase innon-linear oligomers occur at comparable rates. Dextrose is formedrapidly by enzymatic hydrolysis of oligomers, and consumed slowly asnon-linear condensation products are formed, whereas with acid dextroseconcentrations increase slowly.

Optionally, in certain embodiments of the processes described withrespect to U.S. Pat. Nos. 7,608,436, and 8,057,840, enzymatic or acidreversion can be followed by hydrogenation. The hydrogenated productshould have lower caloric content than currently available hydrogenatedstarch hydrolysates. In one embodiment, the hydrogenation can be used todecolorize the product composition without substantially changing itsdextrose equivalence (DE). In one version of the process, enzyme andacid can be used sequentially, in any order. For example, the at leastone catalyst used in the first treatment can be enzyme, and the productcomposition can be subsequently contacted with an acid that acceleratesthe rate of cleavage or formation of glucosyl bonds. Alternatively, theat least one catalyst used in the first treatment can be acid, and theproduct composition can be subsequently contacted with an enzyme thataccelerates the rate of cleavage or formation of glucosyl bonds.

The product composition produced by the treatment with acid, enzyme, orboth, has an increased concentration on a dry solids basis of non-linearsaccharide oligomers. In some cases, the concentration of non-linearsaccharide oligomers having a degree of polymerization of at least three(DP3+) in the product composition is at least about 20%, at least about25%, at least about 30%, or at least about 50% by weight on a dry solidsbasis. In certain such embodiments, the concentration of non-linearsaccharide oligomers having a degree of polymerization of at least three(DP3+) in the product composition is no more than about 100%, or no morethan about 99%, or no more than about 95%, or no more than about 90% byweight on a dry solids basis. In some embodiments, the concentration ofnon-linear saccharide oligomers in the product composition is at leasttwice as high as the concentration of linear saccharide oligomers.

In one particular embodiment of the processes described with respect toU.S. Pat. Nos. 7,608,436, and 8,057,840, the concentration of non-linearsaccharide oligomers in the product composition is at least about 90% byweight on a dry solids basis, and the concentration of isomaltose is atleast about 70% by weight on a dry solids basis.

The product composition will often contain some quantity (typically lessthan 50% by weight on a dry solids basis, and often much less) ofresidual monosaccharides. Optionally, at least some of the residualmonosaccharides (and other species) can be separated from the oligomers(for example by membrane filtration, chromatographic separation, ordigestion via fermentation) and the monosaccharide stream can berecycled into the process feed. In this way, simple sugar syrups can beconverted to high-value food additives.

FIG. 1 shows one embodiment of a process which can make use of thereversion technique described above. The process can begin with astarch, for example a vegetable starch. Conventional corn starch is onesuitable example. The process will generally operate more efficiently ifthe beginning starch has a relatively high purity. In one embodiment,the high purity starch contains less than 0.5% protein on a dry solidsbasis. Although some of the following discussion focuses on corn, itshould be understood that the present invention is also applicable tostarches derived from other sources, such as potato and wheat, amongothers.

Certain embodiments of the processes as described herein with respect toU.S. Pat. Nos. 7,608,436, and 8,057,840 are illustrated schematically inFIG. 9. As shown in FIG. 9, the starch 10 can have acid 12 added to itand can then be gelatinized 14 in a starch cooker, for example in a jetcooker in which starch granules are contacted with steam. In one versionof the process, the starch slurry, adjusted to a pH target of 3.5 byaddition of sulfuric acid, is rapidly mixed with steam in a jet cookerand held at 149 to 152° C. (300 to 305° F.) for 4 minutes in a tailline. The gelatinized starch 16 is hydrolyzed 18 by exposure to acid athigh temperature during jet cooking. The hydrolysis reduces themolecular weight of the starch and generates an increased percentage ofmonosaccharides and oligosaccharides in the composition. (As mentionedabove, the term “oligosaccharides” is used herein to refer tosaccharides comprising at least two saccharide units, for examplesaccharides having a degree of polymerization (DP) of about 2-30.) Aneutralizing agent 20, such as sodium carbonate, can be added to stopthe acid hydrolysis, and then the composition can be furtherdepolymerized 24 by contacting it with a hydrolytic enzyme 22. Suitableenzymes include alpha amylases such as Termamyl, which is available fromNovozymes. This enzymatic hydrolysis further increases the percentage ofmonosaccharides and oligosaccharides present in the composition. Theoverall result of the hydrolysis by acid and enzyme treatment is tosaccharify the starch. The saccharified composition can be isomerized tochange the monosaccharide profile, for example to increase theconcentration of fructose.

The saccharified composition 26 can then be purified, for example bychromatographic fractionation 28. In one embodiment that employs asequential simulated moving bed (SSMB) chromatography procedure, asolution of mixed saccharides is pumped through a column filled withresin beads. Depending on the chemical nature of the resin, some of thesaccharides interact with the resin more strongly leading to a retardedflow through the resin compared to saccharides that interact with theresin more weakly. This fractionation can produce one stream 30 that hasa high content of monosaccharides, such as dextrose and fructose. Highfructose corn syrup is an example of such a stream. The fractionationalso produces a raffinate stream 32 (i.e., faster moving componentsthrough the resin bed) that has a relatively high concentration ofoligosaccharides (e.g., about 5-15% oligosaccharides on a dry solidsbasis (d.s.b.)) and also contains a smaller concentration ofmonosaccharides such as dextrose and fructose. Although the term“stream” is used herein to describe certain parts of the process, itshould be understood that the process of the present invention is notlimited to continuous operation. The process can also be performed inbatch or semi-batch mode.

The raffinate 32 can be further fractionated by membrane filtration 34,for example by nanofiltration, optionally with diafiltration. Forexample, these filtration steps can be performed using a Desal DK spiralwound nanofiltration cartridge at about 500 psi of pressure and at 40-60degrees centigrade temperature. The fractionation described in step 34could also be accomplished by sequential simulated moving bedchromatography (SSMB). The membrane filtration produces a permeate 36(i.e., components that pass through the membrane) which comprisesprimarily monosaccharides, and a retentate 38 (i.e., components rejectedby the membrane) which comprises primarily oligosaccharides.(“Primarily” as used herein means that the composition contains more ofthe listed component than of any other component on a dry solids basis.)The permeate 36 can be combined with the monomer stream 30 (e.g., highfructose corn syrup). The permeate is a monosaccharide-rich stream andthe retentate is an oligosaccharide-rich stream. In other words, thenanofiltration concentrates the oligosaccharides in the retentate andthe monosaccharides in the permeate, relative to the nanofiltrationfeed.

The retentate 38, which can be described as an oligosaccharide syrup 40,can have a sufficiently high content of oligosaccharides that are slowlydigestible (e.g., at least about 50% by weight d.s.b., or in some casesat least about 90%) so that it can be dried or simply evaporated to aconcentrated syrup and used as an ingredient in foods. However, in manycases, it will be useful to further process and purify this composition.Such purification can include one or more of the following steps.(Although FIG. 9 shows four such purification steps 42, 44, 46, and 48as alternatives, it should be understood that two or more of these stepscould be used in the process.)

The oligomers syrup 40 can be subjected to another fractionation 42,such as a membrane filtration, for example a second nanofiltration, inorder to remove at least some of the residual monosaccharides, such asfructose and dextrose. Suitable nanofiltration conditions and equipmentare as described above. This nanofiltration produces a permeate, whichis a second monosaccharide-rich stream, which can be combined with themonomer stream 30. Alternatively, the further fractionation 42 could bedone by chromatographic separation, for example, by simulated mixed-bedchromatography.

The syrup 41 can be isomerized 44 by contacting it with an enzyme suchas dextrose isomerase. This will convert at least some of the residualdextrose present into fructose, which may be more valuable in certainsituations.

The syrup can be treated with an enzyme or acid to cause reversion orrepolymerization 46, in which at least some of the monosaccharides thatare still present are covalently bonded to other monosaccharides or tooligosaccharides, thereby reducing the residual monomer content of thesyrup even further. Suitable enzymes for use in this step includeglucosidases, such as amylase, glucoamylase, transglucosidase, andpullulanase. Cellulase enzymes may produce valuable reversion productsfor some applications.

The syrup can be hydrogenated 48 to convert at least some of anyresidual monosaccharides to the corresponding alcohols (e.g., to convertdextrose to sorbitol). When hydrogenation is included in the process, itwill typically (but not necessarily) be the final purification step.

The purified oligomer syrup 49 produced by one or more of the abovepurification steps can then be decolorized 50. Decolorization can bedone by treatment with activated carbon followed by microfiltration, forexample. In continuous flow systems, syrup streams can be pumped throughcolumns filled with granular activated carbon to achieve decolorization.The decolorized oligomer syrup can then be evaporated 52, for example toabout greater than about 70% dry solids (d.s.), giving a product thatcomprises a high content of oligosaccharides (e.g., greater than 90% bywt d.s.b., and in some instances greater than 95%), and acorrespondingly low monosaccharide content. The product comprises aplurality of saccharides which are slowly or incompletely digested byhumans, if not totally indigestible. These sugars can includeisomaltose, panose and branched oligomers having a degree ofpolymerization of four or greater.

The process conditions can be modified to recover the majority of themaltose in the feed either in the monomer-rich streams (30, 36) or inthe oligomer product stream. For example, a nanofiltration membrane witha slightly larger pores, such as Desal DL, operating at less than 500psi pressure can be used to increase the amount of maltose inmonomer-rich streams.

In certain embodiments of the methods and compositions as describedherein, the fermentable soluble fiber is a slowly digestible saccharideoligomer composition that is suitable for use in foods. “Slowlydigestible” as the term is used herein means that one or morecarbohydrates are either not digested at all in the human stomach andsmall intestine, or are only digested to a limited extent. Both in vitroand in vivo tests can be performed to estimate the rate and extent ofcarbohydrate digestion in humans. The “Englyst Assay” is an in vitroenzyme test that can be used to estimate the amounts of a carbohydrateingredient that are rapidly digestible, slowly digestible or resistantto digestion (European Journal of Clinical Nutrition (1992) Volume 46(Suppl. 2), pages S33-S50). Thus, any reference herein to “at leastabout 50% by weight on a dry solids basis” of a material being “slowlydigestible” means that the sum of the percentages of that material thatare classified as slowly digestible or as resistant by the Englyst assaytotals at least about 50%. The terms “oligosaccharides” and “saccharideoligomers” are used herein to refer to saccharides comprising at leasttwo saccharide units, for example saccharides having a degree ofpolymerization (“DP”) of about 2-30. For example, a disaccharide has aDP of 2.

Gastrointestinal enzymes readily recognize and digest carbohydrates inwhich the dextrose units are linked alpha (1->4) (“linear” linkages).Replacing these linkages with alternative linkages (alpha (1->3), alpha(1->6) (“non-linear” linkages) or beta linkages, for example) greatlyreduces the ability of gastrointestinal enzymes to digest thecarbohydrate. This will allow the carbohydrates to pass on into thesmall intestines largely unchanged. In certain embodiments of themethods and compositions as described herein, the fermentable solublefiber, e.g., the soluble corn fiber, comprises a minor amount (i.e.,less than 50 wt % on a dry solids basis, and usually a much lowerconcentration, e.g., less than 40 wt %, less than 30 wt %) of residualmonosaccharides. In some embodiments as described herein, at least about50% by weight on a dry solids basis of the product composition is slowlydigestible. The processes as described with respect to U.S. Pat. Nos.7,608,436, and 8,057,840 can include the additional step of removing atleast some of the residual monosaccharides (and optionally other speciesas well) from the product composition by membrane filtration,chromatographic fractionation, or digestion via fermentation. Theseparated monosaccharides can be combined with other process streams,for example for production of dextrose or corn syrup. Alternatively, theseparated monosaccharides can be recycled into the feed composition.

In certain embodiments of the methods and compositions as describedherein, the fermentable soluble fiber comprises a major amount (e.g.,greater than 50%, greater than about 60%, or greater than about 70%) ona dry solids basis of linear and non-linear saccharide oligomers, andwherein the concentration of non-linear saccharide oligomers is greaterthan the concentration of linear saccharide oligomers, and wherein theconcentration of non-linear saccharide oligomers having a degree ofpolymerization of at least three is at least about 20% by weight on adry solids basis. For example, in certain embodiments, the concentrationof non-linear saccharide oligomers in the composition is at least twiceas high as the concentration of linear saccharide oligomers. In certainembodiments, the concentration of non-linear saccharide oligomers havinga degree of polymerization of at least three is at least about 25% byweight on a dry solids basis. In certain embodiments, the concentrationof non-linear saccharide oligomers having a degree of polymerization ofat least three is at least about 30% by weight, or even at least 50% byweight, on a dry solids basis. In certain embodiments, wherein theconcentration of non-linear saccharide oligomers is at least about 90%by weight on a dry solids basis, and the concentration of isomaltose isat least about 70% by weight on a dry solids basis.

In certain embodiments of the methods and compositions described herein,the fermentable soluble fiber, e.g., soluble corn fiber, is administeredat a rate of at least about 3 g/day. For example, in certainembodiments, the fermentable soluble fiber, e.g., soluble corn fiber, isadministered at a rate of at least about 5 g/day, at least about 7g/day, at least about 10/day, at least about 12 g/day, at least about 13g/day, at least about 15 g/day, or even at least about 20 g/day and nomore than about 100 g/day, or no more than 75 g/day. Specifically,clinically relevant gastrointestinal tolerance has been established at65 g/day when spread over 12 hrs (a normal eating day) and/or at 40g/acute bolus day. These are both well tolerated doses. Accordingly, incertain such embodiments, the fermentable soluble fiber, e.g., solublecorn fiber, is administered at a rate of no more than about 65 g over 12hours, and/or no more than about 40 g in a single bolus.

For example, in certain embodiments of the methods and compositionsdescribed herein, the fermentable soluble fiber, e.g., soluble cornfiber, is administered at a rate in the range of about 3 g/day to about100 g/day. In other embodiments of the methods and compositionsdescribed herein, the fermentable soluble fiber, e.g., soluble cornfiber, is administered at a rate in the range of about 10 g/day to about100 g/day, or about 12 g/day to about 100 g/day. In other embodiments ofthe methods and compositions described herein, the fermentable solublefiber, e.g., soluble corn fiber, is administered at a rate in the rangeof about 5 to about 65 g/day, about 5 to about 40 g/day, about 5 toabout 30 g/day, about 5 to about 20 g/day, about 10 to about 65 g/day,about 10 to about 40 g/day, about 10 to about 30 g/day, about 15 toabout 65 g/day, about 15 to about 40 g/day, about 15 to about 30 g/day,about 5 to about 15 g/day, about 7 to about 15 g/day, about 9 to about15 g/day, or about 10 to about 15 g/day, about 12 to about 20 g/day,about 13 to about 20 g/day, about 14 to about 20 g/day, about 15 toabout 20 g/day, about 16 to about 20 g/day, about 17 to about 20 g/day,about 18 to about 20 g/day, or about 19 to about 20 g/day. In certainembodiments of the methods and compositions described herein, thefermentable soluble fiber, e.g., soluble corn fiber, is administered ata rate of about 5 g/day, about 6 g/day, about 7 g/day, about 8 g/day,about 9 g/day, or about 10 g/day. In other embodiments of the methodsand compositions described herein, the fermentable soluble fiber, e.g.,soluble corn fiber, is administered at a rate in the range of about 11to about 20 g/day. In other embodiments of the methods and compositionsdescribed herein, the fermentable soluble fiber, e.g., soluble cornfiber, is administered at a rate of about 11 g/day, or about 12 g/day,or about 13 g/day, or about 14 g/day, or about 15 g/day, or about 16g/day, or about 17 g/day, or about 18 g/day, or about 19 g/day, or about20 g/day.

In a given day, the administration can be broken up into any number ofdosages. For example, in one embodiment of the methods and compositionsdescribed herein, the fermentable soluble fiber, e.g., soluble cornfiber, is administered once per day (e.g., in a single serving). Inother embodiments of the methods and compositions described herein, thefermentable soluble fiber, e.g., soluble corn fiber, is administered aplurality of times a day, for example, twice per day or three times perday (e.g., in a plurality of servings, for example, in two servings orin three servings per day). When a plurality of administrations orservings is to be used, the amounts per day described above can bedivided among the number of administrations or servings to provideacceptable amounts per serving that are well tolerated (i.e., does notcause severe bloating, flatulence, stomach noises, abdominal cramps,diarrhea, nausea, and/or vomiting).

In another aspect, the disclosure provides a method of increasingmineral (e.g., calcium, iron, zinc, copper, potassium and/or magnesium)absorption in a subject, where the method includes orally administeringto the subject a fermentable soluble fiber, e.g., soluble corn fiber. Incertain such embodiments, the mineral for which absorption is increasedis a mineral, such as, for example calcium and/or iron. In certainembodiments of the methods and compositions as described herein, themineral is absorbed as a divalent cation. In certain embodiments of themethods and compositions as described herein, the mineral is calcium. Inother embodiments of the methods and compositions as described herein,the mineral is calcium and/or magnesium. In certain embodiments of themethods and compositions as described herein, the mineral is calciumand/or iron. In other embodiments of the methods and compositions asdescribed herein, the mineral is calcium, magnesium and/or iron. Theadministration can, in certain embodiments, be as otherwise describedherein.

In another embodiment, the disclosure provides a method of increasingone or more colonic bacteria populations and increasing mineral (e.g.,calcium, iron, zinc, copper, potassium and/or magnesium) absorption in asubject, where the method includes orally administering to a fermentablesoluble fiber, e.g., soluble corn fiber. The administration can, incertain embodiments, be as otherwise described herein.

In certain embodiments of the methods and compositions described herein,calcium absorption is increased by at least about 3% as compared to anon-treated subject. In certain embodiments of the methods andcompositions described herein, calcium absorption is increased by atleast about 5%, at least about 6%, at least about 7%, at least about 8%,at least about 9%, at least about 10%, at least about 11%, at leastabout 12%, at least about 13%, or at least about 14%, or at least about15%, as compared to the non-treated subject. In other embodiments of themethods and compositions described herein, calcium absorption isincreased by at least about 20%, or at least about 25% as compared to anon-treated subject. In certain embodiments of the methods andcompositions described herein, calcium absorption is increased by atleast about 20%, at least about 25%, at least about 30%, or at leastabout 35% as compared to a non-treated subject. In certain suchembodiments, calcium absorption is increased by no more than about 200%as compared to a non-treated subject. In other such embodiments, calciumabsorption is increased by no more than about 100% as compared to anon-treated subject. In other such embodiments, calcium absorption isincreased by no more than about 50% as compared to a non-treatedsubject. The timing for calcium absorption can be, for example, in therange of 24-48 h, e.g., at 36 h or 48 h.

In certain embodiments of the methods and compositions described herein,the subject is a mammal. In one embodiment of the methods andcompositions described herein, the subject is a human, for example, anon-adult human (e.g., in the range of about 2 years old to about 20years old, or about 13 years old to about 19 years old), or an olderhuman (e.g., at least about 45 years old, at least about 50 years old,at least about 60 years old, at least about 70 years old, at least about80 years old or even at least about 90 years old, especially an olderfemale human). Accordingly, in certain embodiments the methods andcompositions described herein can be used with subjects who areespecially likely to benefit from increased mineral (e.g., calcium)absorption.

It is envisioned that the effects of increasing one or more colonicbacteria populations and/or increasing calcium absorption relate to bothhumans and animals, and thus can be applied to foodstuffs and animalfeed. Representative non-human animals include, livestock, such ashorses, chicken, turkeys, cattle, cow, swine, sheep, goats, llamas andbison, cats and dogs, rodents, rabbits, hamsters and birds.

The administration can be performed over an extended time period, forexample, over the course of at least about a week, over the course of atleast about two weeks, of at least about three weeks over the course ofat least about four weeks, of at least about seven weeks or even overthe course of at least about 52 weeks. The person of ordinary skill inthe art that in such long-term administrations, days of administrationmay be “missed”; desirably the number of days missed is less than about10% of the total number of days over which the administration isperformed.

Another embodiment of the invention is an edible composition thatincludes at least about 2.5 g of fermentable soluble fiber, e.g.,soluble corn fiber, per serving. For example, certain embodiments ofedible compositions as described herein include at least about 3 g, atleast about 4 g, at least about 5 g, at least about 6 g, at least about8 g, at least about 10 g, or even at least about 20 g of fermentablesoluble fiber, e.g., soluble corn fiber, per serving. In certain suchembodiments, the edible composition includes no more than 100 g, no morethan about 50 g, or even no more than about 40 g of fermentable solublefiber, e.g., soluble corn fiber, per serving. The edible compositionscan, for example, be provided as food compositions as described below.In other embodiments, an edible composition is provided as a nutritionalsupplement. Such edible compositions can be useful in performing themethods described herein.

In certain such embodiments of the compositions as described herein, thesize of the serving can be, for example, at least about 75 g, at leastabout 150 g, or even at least about 200 g. In certain embodiments, thesize of the serving is no more than about 1000 g, or even no more thanabout 500 g. For example, in one embodiment, the serving size in therange of about 75 mL to about 1000 mL. In certain embodiments, eachserving is separately packaged. In other embodiments, multiple servingsare packaged together, and provided with instructions relating a servingsize and/or an amount of fermentable soluble fiber, e.g., soluble cornfiber, per serving as described herein.

Another embodiment of the invention is an edible composition thatincludes fermentable soluble fiber, e.g., soluble corn fiber, in anamount of at least about 2.5%, at least about 3%, at least about 5%, atleast about 10%, at least about 20%, at least about 30%, or even atleast about 40% by weight. However, in certain such embodiments, theedible composition has a maximum amount of fermentable soluble fiber,e.g., soluble corn fiber, that is no greater than about 75%, or even nogreater than about 50% by weight. The edible compositions can, forexample, be provided as food compositions as described below. The ediblecompositions can, for example, be provided with the serving sizes and/orthe amounts of soluble corn fiber per serving as described herein.

Another embodiment of the invention is an edible composition thatincludes one or more (e.g., two or more, or three or more) bacterialpopulations, each from a genus selected from the group consisting ofLactobacillus, Bacteroides, Parabacteroides, Alistipes, Bifidobacterium,Butyricicoccus, Oscillibacter, and Dialister, as well as the bacteriaindicated as increasing in population with soluble corn fiberadministration in Table 5, and the bacteria indicated as beingcorrelated with calcium absorption in Table 6. One or more of thebacterial populations, can, for example, act as probiotics. The ediblecompositions described herein can, in certain embodiments, includefermentable soluble fiber, e.g., soluble corn fiber, (for example, in anamount as described above). But in other embodiments, the ediblecomposition does not include a fermentable soluble fiber. Suchembodiments can be useful, for example, for addition to orco-administration with compositions including fermentable solublefibers, such that the bacterial populations of the edible compositionare present in the colon at the same time as the fermentable solublefiber. Accordingly, the subject can in certain embodiments enjoy thebenefits of the combination of fermentable soluble fibers with thebacterial populations identified herein without being administered asingle composition that includes both the fermentable soluble fibers andthe bacterial populations. Similarly, products suitable for theenjoyment the benefits of the combination of the fermentable solublefibers with the bacterial populations identified herein can beformulated that do not include both the fermentable soluble fiber andthe bacterial populations.

For example, in certain embodiments of the edible compositions describedherein, the edible composition includes one or more (e.g., two or more,or three or more) bacteria populations, each from a genus (e.g., eachfrom a different genus) selected from the group consisting ofParabacteroides, Butyricicoccus, Oscillibacter, and Dialister, and anycombination thereof. For example, one embodiment of the ediblecompositions described herein includes a population of Parabacteroides.Another embodiment of the edible compositions described herein includesa population of Butyricicoccus. Another embodiment of the ediblecompositions described herein includes a population of Oscillibacter.Another embodiment of the edible compositions described herein includesa population of Dialister. For example, certain embodiments of theedible compositions described herein include populations ofParabacteroides and Butyricicoccus; Parabacteroides and Oscillibacter,Parabacteroides and Dialister; Butyricicoccus and Oscillibacter;Butyricicoccus and Dialister, Oscillibacter and Dialister;Parabacteroides, Butyricicoccus and Oscillibacter, Parabacteroides,Butyricicoccus and Dialister, Parabacteroides, Oscillibacter, andDialister, Butyricicoccus, Oscillibacter, and Dialister, orParabacteroides, Butyricicoccus, Oscillibacter, and Dialister.

In other embodiments of the edible compositions described herein, theedible composition includes one or more (e.g., two or more, or three ormore) bacteria populations, each from a genus (e.g., each from adifferent genus) selected from the group consisting of Bacteroides,Butyricicoccus, Oscillibacter, and Dialister, and any combinationthereof. For example, one embodiment of the edible compositionsdescribed herein includes a population of Bacteroides. Anotherembodiment of the edible compositions described herein includes apopulation of Butyricicoccus. Another embodiment of the ediblecompositions described herein includes a population of Oscillibacter.Another embodiment of the edible compositions described herein includesa population of Dialister. For example, certain embodiments of theedible compositions described herein include populations of Bacteroidesand Butyricicoccus; Bacteroides and Oscillibacter, Bacteroides andDialister; Butyricicoccus and Oscillibacter, Butyricicoccus andDialister, Oscillibacter and Dialister, Bacteroides, Butyricicoccus andOscillibacter; Bacteroides, Butyricicoccus and Dialister;Parabacteroides, Oscillibacter, and Dialister, Butyricicoccus,Oscillibacter, and Dialister, or Parabacteroides, Butyricicoccus,Oscillibacter, and Dialister.

In other embodiments of the edible compositions described herein, theedible composition includes one or more (e.g., two or more, or three ormore) bacteria populations, each from a genus (e.g., each from adifferent genus) selected from the group consisting of Parabacteroides,Bifidobacterium, Alistipes, Anaerococcus, Catenibacterium, genera withinthe order Clostridiales (e.g., not Clostridium, Anaerofustis,Anaerococcus, Coprococcus, Peptostreptococcaceae, Sporacetigenium); andgenera within the family Ruminococcaceae and any combination thereof.For example, one embodiment of the edible compositions described hereinincludes a population of Parabacteroides. Another embodiment of theedible compositions described herein includes a population ofBifidobacterium. Another embodiment of the edible compositions describedherein includes a population of Alistipes. Another embodiment of theedible compositions described herein includes a population ofAnaerococcus. Another embodiment of the edible compositions describedherein includes a population of Catenibacterium. Another embodiment ofthe edible compositions described herein includes a population ofRuminococcaceae. Another embodiment of the edible compositions describedherein includes a population of Clostridiales. For example, certainembodiments of the edible compositions described herein includepopulations of Parabacteroides and Bifidobacterium; Parabacteroides andAlistipes; Parabacteroides and Anaerococcus; Parabacteroides andCatenibacterium; Parabacteroides and Ruminococcaceae; Parabacteroidesand Clostridiales; Bifidobacterium and Alistipes; Bifidobacterium andAnaerococcus; Bifidobacterium and Catenibacterium; Bifidobacterium andRuminococcaceae; Bifidobacterium and Clostridiales; Alistipes andAnaerococcus; Alistipes and Catenibacterium; Alistipes andRuminococcaceae; Anaerococcus and Catenibacterium; Anaerococcus andRuminococcaceae; Anaerococcus and Clostridiales; Catenibacterium andRuminococcaceae; Catenibacterium and Clostridiales; Ruminococcaceae andClostridiales; Parabacteroides, Bifidobacterium and Alistipes;Parabacteroides, Bifidobacterium and Anaerococcus; Parabacteroides,Bifidobacterium and Catenibacterium; Parabacteroides, Bifidobacteriumand Clostridiales; Parabacteroides, Bifidobacterium and Ruminococcaceae;Parabacteroides, Alistipes and Anaerococcus; Parabacteroides, Alistipesand Catenibacterium; Parabacteroides, Alistipes and Clostridiales;Parabacteroides, Alistipes and Ruminococcaceae; Parabacteroides,Anaerococcus and Catenibacterium; Parabacteroides, Anaerococcus andClostridiales; Parabacteroides, Anaerococcus and Ruminococcaceae;Parabacteroides, Catenibacterium and Clostridiales; Parabacteroides,Catenibacterium and Ruminococcaceae; Parabacteroides, Clostridiales andRuminococcaceae; Bifidobacterium, Alistipes and Anaerococcus;Bifidobacterium, Alistipes and Catenibacterium; Bifidobacterium,Alistipes and Clostridiales; Bifidobacterium, Alistipes andRuminococcaceae; Bifidobacterium, Anaerococcus and Catenibacterium;Bifidobacterium, Anaerococcus and Clostridiales; Bifidobacterium,Anaerococcus and Ruminococcaceae; Bifidobacterium, Catenibacterium andClostridiales; Bifidobacterium, Catenibacterium and Ruminococcaceae;Bifidobacterium, Clostridiales and Ruminococcaceae; Alistipes,Anaerococcus and Catenibacterium; Alistipes, Anaerococcus andClostridiales; Alistipes, Anaerococcus and Ruminococcaceae; Alistipes,Catenibacterium and Clostridiales; Alistipes, Catenibacterium andRuminococcaceae; Alistipes, Clostridiales and Ruminococcaceae;Anaerococcus, Catenibacterium and Clostridiales; Anaerococcus,Catenibacterium and Ruminococcaceae; Anaerococcus, Clostridiales andRuminococcaceae; Catenibacterium, Clostridiales and Ruminococcaceae;Parabacteroides, Bifidobacterium, Alistipes and Anaerococcus;Parabacteroides, Bifidobacterium, Alistipes and Catenibacterium;Parabacteroides, Bifidobacterium, Alistipes and Clostridiales;Parabacteroides, Bifidobacterium, Alistipes and Ruminococcaceae;Parabacteroides, Bifidobacterium, Anaerococcus and Catenibacterium;Parabacteroides, Bifidobacterium, Anaerococcus and Clostridiales;Parabacteroides, Bifidobacterium, Anaerococcus and Ruminococcaceae;Parabacteroides, Bifidobacterium, Catenibacterium and Clostridiales;Parabacteroides, Bifidobacterium, Catenibacterium and Ruminococcaceae;Parabacteroides, Bifidobacterium, Clostridiales and Ruminococcaceae;Parabacteroides, Alistipes, Anaerococcus and Catenibacterium;Parabacteroides, Alistipes, Anaerococcus and Clostridiales;Parabacteroides, Alistipes, Anaerococcus and Ruminococcaceae;Parabacteroides, Alistipes, Catenibacterium and Clostridiales;Parabacteroides, Alistipes, Catenibacterium and Ruminococcaceae;Parabacteroides, Alistipes, Clostridiales and Ruminococcaceae;Parabacteroides, Anaerococcus, Catenibacterium and Clostridiales;Parabacteroides, Anaerococcus, Catenibacterium and Ruminococcaceae;Parabacteroides, Anaerococcus, Clostridiales and Ruminococcaceae;Parabacteroides, Catenibacterium, Clostridiales and Ruminococcaceae;Bifidobacterium, Alistipes, Anaerococcus and Catenibacterium;Bifidobacterium, Alistipes, Anaerococcus and Clostridiales;Bifidobacterium, Alistipes, Anaerococcus and Ruminococcaceae;Bifidobacterium, Alistipes, Catenibacterium and Clostridiales;Bifidobacterium, Alistipes, Catenibacterium and Ruminococcaceae;Bifidobacterium, Alistipes, Clostridiales and Ruminococcaceae;Bifidobacterium, Anaerococcus, Catenibacterium and Clostridiales;Bifidobacterium, Anaerococcus, Catenibacterium and Ruminococcaceae;Bifidobacterium, Anaerococcus, Clostridiales and Ruminococcaceae;Bifidobacterium, Catenibacterium, Clostridiales and Ruminococcaceae;Alistipes, Anaerococcus, Catenibacterium and Clostridiales; Alistipes,Anaerococcus, Catenibacterium and Ruminococcaceae; Alistipes,Anaerococcus, Clostridiales and Ruminococcaceae; Alistipes,Catenibacterium, Clostridiales and Ruminococcaceae; or Anaerococcus,Catenibacterium, Clostridiales and Ruminococcaceae. Of course, theperson of ordinary skill in the art will appreciate that any combinationof 5, 6, or 7 colonic bacteria populations, each from a different genusselected from the group consisting of Parabacteroides, Bifidobacterium,Alistipes, Anaerococcus, Catenibacterium, genera within the orderClostridiales; and genera within the family Ruminococcaceae, may beincluded in the edible compositions described herein.

In other embodiments of the edible compositions described herein, theedible composition includes one or more (e.g., two or more, or three ormore) bacteria populations, each from a genus (e.g., each from adifferent genus) selected from the group consisting of Parabacteroides,Dialister, Akkermansia, and genera within the family Lachnospiraceae(e.g., not Lachnospira). For example, one embodiment of the ediblecompositions described herein includes a population of Parabacteroides.Another embodiment of the edible compositions described herein includesa population of Dialister. Another embodiment of the edible compositionsdescribed herein includes a population of Akkermansia. Anotherembodiment of the edible compositions described herein includes apopulation of Lachnospiraceae. For example, certain embodiments of theedible compositions described herein include populations ofParabacteroides and Dialister, Parabacteroides and Akkermansia;Parabacteroides and Lachnospiraceae; Dialister and Akkermansia;Dialister and Lachnospiraceae; Akkermansia and Lachnospiraceae;Parabacteroides, Dialister, and Akkermansia; Parabacteroides, Dialister,and Lachnospiraceae; Parabacteroides, Akkermansia, and Lachnospiraceae;Dialister, Akkermansia, and Lachnospiraceae; or Parabacteroides,Dialister, Akkermansia, and Lachnospiraceae.

Of course, as the person of ordinary skill in the art will appreciate,the edible compositions including specific combinations of bacteriapopulations as described herein can further include other bacteriapopulations, either as elsewhere described herein or otherwise. Forexample, the compositions can further include one or more bacteriapopulations selected from the genera Bifidobacterium and Lactobacillus.

The edible compositions can, for example, be provided as foodcompositions as described below. In other embodiments, an ediblecomposition is provided as a nutritional supplement. In still otherembodiments, an edible composition is provided as an ingredient to bemixed with a food composition, for example, during processing orcooking, or at the time of serving or eating. The edible compositionscan, for example, be provided with the fermentable soluble fiber, e.g.,soluble corn fiber, concentrations, the serving sizes and/or the amountsof fermentable soluble fiber, e.g., soluble corn fiber, per serving asdescribed herein. The amount of the bacterial populations added to thecomposition may be adjusted by the person of skill in the art to meetthe desired need. In general, each of the bacterial populations may bein the amount of about 1×10³ to about 1×10¹⁰ CFU (colony-forming unit).In certain embodiments, each of the bacterial populations is in theamount of about 1×10⁵ to about 1×10¹⁰ CFU, or about 1×10⁶ to about1×10¹⁰ CFU, or about 1×10⁷ to about 1×10¹⁰ CFU, or about 1×10⁸ to about1×10¹⁰ CFU, or about 1×10³ to about 1×10⁸ CFU, or about 1×10⁴ to about1×10⁸ CFU, or about 1×10⁵ to about 1×10⁸ CFU, or about 1×10⁶ to about1×10⁸ CFU, or about 1×10⁵ to about 1×10⁷ CFU, or about 1×10⁴ CFU, orabout 1×10⁵ CFU, or about 1×10⁶ CFU, or about 1×10⁷ CFU or about 1×10⁸CFU, or about 1×10⁹ CFU, or about 1×10¹⁰ CFU.

Another embodiment of the invention is an edible composition asdescribed above that further includes one or more mineral species. Eachmineral species can, for example, be a divalent mineral species, or aspecies selected from a calcium species, a magnesium species, a copperspecies, a potassium species, a zinc species and an iron species. Forexample, in one embodiment, the edible composition includes calcium. Inanother embodiment, the edible composition includes calcium and/ormagnesium. In another embodiment, the edible composition includescalcium, magnesium, and/or iron. The mineral species can be provided,for example, as a salt, such as a carbonate salt, a halide salt, or abicarbonate salt. Calcium, for example, can be provided as, e.g.,calcium carbonate or calcium gluconate. The mineral (e.g., the calcium)can be provided, for example, at an amount of at least about 50 mg perdose or serving, at least about 100 mg per dose or serving, at leastabout 250 mg per dose or serving, at least about 500 mg per dose orserving, or even at least about 1000 mg per dose or serving. In certainsuch embodiments, the calcium is included at less than about 2000 mg perdose or serving or even less than about 1000 mg per dose or serving. Theedible compositions can, for example, be provided as food compositionsas described below. In other embodiments, an edible composition isprovided as a nutritional supplement. The edible compositions can, forexample, be provided with the fermentable soluble fiber, e.g., solublecorn fiber, concentrations, the serving sizes and/or the amounts offermentable soluble fiber, e.g., soluble corn fiber, per serving asdescribed herein.

In other embodiments, the composition of the disclosure does not includea mineral species as described above.

Another embodiment of the invention is an edible composition asdescribed above that further includes one or more additional prebiotics.Examples of prebiotics include, but are not limited to, inulin,lactulose, fructooligosaccharide, mannooligosaccharide, larcharabinogalactan, xylooligosaccharide, polydextrose, and tagatose. Incertain embodiments, the disclosure provides edible compositions asdescribed above, wherein the prebiotic is in the range of 0.025 g to 10g. In certain embodiments, the prebiotic is in the amount of about 0.1to about 10 g, or about 1 to about 10 g, or about 0.1 to about 5 g, orabout 1 to about 5 g, or about 5 to about 10 g, or about 5 to about 8 g,or about 2 to about 8 g, or about 2 to about 5 g, or about 2 to about 8g, or about 0.05 g, or about 0.1 g, or about 1 g, or about 2 g or about5 g, or about 8 g, or about 10 g.

In one embodiment, the composition of the disclosure does not includeone or more additional prebiotics as described above. For example, inone embodiment, the compositions of the disclosure do not include one ormore of the prebiotics selected from the group consisting of inulin,lactulose, fructooligosaccharide, mannooligosaccharide, larcharabinogalactan, xylooligosaccharide, polydextrose, and tagatose. Inanother embodiment, the compositions of the disclosure does not includeinulin. In yet another embodiment, the compositions of the disclosuredoes not include pullulan.

Optionally, an edible composition or a food composition can also includeadditional nutritive or non-nutritive saccharides and/orpolysaccharides. In one embodiment, the edible composition comprisessorbitol, pullulan, or a combination thereof. Sorbitol delivers about60% of the sweetness of sugar to foods, but at a significant reductionin caloric content (2.6 vs. 4.0 kcal/g, Livesay) and with a negligibleglycemic response. Pullulan gum is a slowly digestible carbohydrate thatgives about a 50% relative glycemic response in humans compared torapidly digestible carbohydrate, but may deliver similar caloric contentas sugar to foods.

In one embodiment, the food product comprises about 50-99% fermentablesoluble fiber, e.g., soluble corn fiber, 0-50% fructose, 0-33% pullulan,and 0-33% sorbitol, provided that the concentration of at least one offructose, pullulan, or sorbitol is at least 1%. (All of thesepercentages are by weight.) In another embodiment, the food productcomprises about 60-80% fermentable soluble fiber, e.g., soluble cornfiber, 1-20% fructose, 0-20% pullulan, and 0-20/o sorbitol. In yetanother embodiment, the food product comprises about 65-75% fermentablesoluble fiber, e.g., soluble corn fiber, 5-15% fructose, 5-15% pullulan,and 5-15% sorbitol. In embodiments that comprise a high intensitysweetener, the concentration of that ingredient can be about 0.001-0.5%.

The edible composition or food composition optionally can also containresistant starch or other fiber sources.

As the person of skill in the art will appreciate, the compositionsdescribed herein can be used in practicing the methods describedelsewhere herein.

The terms “edible” and “edible composition” are used in a broad senseherein to include a variety of substances that can be ingested byhumans, such as food, beverages and medicinal and nutritional supplementdosage forms such as syrups, powders, capsules or tablets. The terms“food” and “food composition” are used more narrowly to mean foods andbeverages and ingredients therefor. Suitable food compositions can be ina variety of forms including, but are not limited to baked foods,breakfast cereal, dairy products, soy products, confections, jams andjellies, beverages (powdered and/or liquid), shakes, fillings, yogurts(dairy and non-dairy yogurts), kefirs, extruded and sheeted snacks,gelatin desserts, snack bars, meal replacement and energy bars, cheeseand cheese sauces (dairy and non-dairy cheeses), edible andwater-soluble films, soups, syrups, table top sweeteners, nutritionalsupplements, sauces, dressings, creamers, icings, ice cream, frostings,glazes, pet food, tortillas, meat and fish, dried fruit, infant andtoddler food, and batters and breadings.

In order to make the food product suitable for use as a flavor enhancerin food compositions, in many cases it will be desirable for it also toinclude a natural and artificial flavors. Suitable examples of suchflavors include apple, citrus, grape, orange, cherry, lemon, lime,vanilla, peach, peanut butter, pineapple, pomegranate, blueberry,raspberry, blackberry, jasmine, lavender, mint, strawberry, banana,mango, passion fruit, dragon fruit, kiwi, chocolate, maple, rum, butter,and combinations thereof.

In certain embodiments, the edible composition is in the form of anagglomerated powder, for example, like those used in making powdereddrinks and nutritional supplements.

In order to make the food product suitable for use as a sweetenercomposition in food, in many cases it will be desirable for it also toinclude a non-nutritive high-intensity sweetener. Suitable examples ofsuch non-nutritive high-intensity sweeteners include, but are notlimited to sucralose, acesulfame potassium, aspartame, monkfruit,Stevia, and combinations thereof.

The person of ordinary skill in the art will appreciate that thefermentable soluble fiber, e.g., soluble corn fiber, can be provided inany of several different physical forms, such as powder, agglomeratedpowder, syrup or concentrated syrup solids. In one embodiment, thesoluble fiber is in particulate form. The particulates can be heldtogether by a binder, such as a binder composition that comprises amajor amount of maltodextrin. An agglomeration of particulates can haveadvantages in terms of rate of dissolution and dispersion. This can beuseful in applications where more rapid dissolution and lower shearrates of mixing are important, such as table top sugar replacement,table top fiber supplementation, and on-the-go dry powder drink mixproducts.

Additional aspects suitable for use in edible compositions as describedherein are described further in U.S. Patent Applications Publicationsnos. 2008/0292766, 2006/0210696 and 2008/0175977, each of which ishereby incorporated herein by reference in its entirety, and which isattached in the appendix to this specification. In certain embodimentsof the methods and compositions described herein, the edible compositionis in a form and uses additional ingredients as described in an aspector embodiment of U.S. Patent Application Publication no. 2008/0292766,2006/0210696 or 2008/0175977.

Certain aspects of the invention are further described with respect tothe experimental studies described below.

Example 1 Subjects and Methods Subjects

Fifteen boys, aged 13-15 y, and 9 girls, aged 12-14 y, participated inthese metabolic studies. Screening questionnaires were used to determineeligibility based on a brief medical history, maturational age, physicalactivity, and habitual dietary intake assessed with a 6-day diet record.Exclusion criteria included abnormal liver or kidney function,malabsorptive disorders, anemia, smoking, history of medications thatinfluence calcium metabolism (steroids, thiazide diuretics), body weightoutside the 5-95^(th) BMI percentile for age, regular consumption ofillegal drugs, non-prescription drugs, or any kind of contraceptives,and pregnancy. Subjects were not permitted to take nutritionalsupplements while participating in these studies and were asked todiscontinue use before coming to camp.

Study Design

This study, designed to havoc a summer camp environment, was composed oftwo 3-week balance studies separated by a 7-day washout period. Thistrial used a double-blind, cross-over design in which participantsreceived two treatments in randomized order, 12 g soluble corn fiber orplacebo.

Diets

Controlled diets were provided throughout both camp sessions andcontained foods that are typically eaten by adolescent children such asspaghetti, hamburgers, sandwiches and potato chips. Subjects wereassigned to one of five energy levels (1750, 2100, 2400, 2700, and 3000kilocalories) based on estimated energy requirements calculated usingthe Harris-Benedict equations. Diets were designed to maintain bodyweight and to contain constant levels of key nutrients. The controlleddiet was provided as a 4-day cycle menu with 3 meals and 2 snacks daily.On average the diet contained 14% protein, 33% fat, 53% carbohydrate,200 IU vitamin D, 1100 mg phosphorus, 2300 mg sodium, and 600 mgcalcium. Fifteen grams of fiber was included in the basal diet and theintervention added an additional 0 or 12 g of SCF. This yielded a totaldietary fiber content of 15 and 27 g for the Control and SCF treatments,respectively. SCF was given in WELCH'S® fruit snacks and divided intotwo 0 or 6 g doses provided at lunch and dinner. SCF provided by Tate &Lyle Health & Nutrition Sciences (Hoffman Estates, Ill.), contained >70%soluble dietary fiber with an approximate weight-average degree ofpolymerization of 10 and α-1,4, α-1,6, α-1,3, and α-1,2 bonds.

Anthropometrics and Bone Measurements

Anthropometric measures including weight, sitting height, bitrochantericwidth, waist circumference, and hip circumference were taken during thefirst session of camp. Standing height using a wall-mounted stadiometerwas measured at the beginning of the first session and weight wasmonitored in the morning on each day with an electronic digital scale toensure that weight remained stable throughout the sessions. Bone mineralcontent (BMC) and bone mineral density (BMD) were measured by dualenergy x-ray absorptiometry (DXA) (GE Lunar, Madison, Wis.) during onebalance period. Bone measurements were taken of the total body, spine,forearm, and both hips.

Hormones and Biochemical Markers of Bone Metabolism

A fasting, baseline blood draw was taken on the first day of camp fordetermination of general blood chemistries to verify clinical profilesand health of the participants. A second fasting sample was taken at theend of camp to measure biochemical markers of bone dynamics and hormonesrelated to calcium and vitamin D metabolism.

Sample Collection and Analysis

All urine and fecal samples were collected from day 1 to 21 of eachbalance period and pooled as 24-h collections. Calcium content of diet,fecal, and urine samples was measured using inductively coupled plasmaoptical emission spectrometry (Optima 4300 DV, Perkin Elmer Instrument)as previously described. All fecal samples were frozen and processed ata later time for calcium. Urine was refrigerated and also analyzed laterfor total calcium content.

Compliance

Participants were supervised during activities, meals, and samplecollection by trained counselors for 24 h each day. Unconsumed food frommeals was collected and recorded. Urine collection compliance wasevaluated by measuring creatinine excreted in urine by enzymaticcolorimetric assay (COBAS Integra, Roche Diagnostics). Fecal collectioncompliance was assessed by polyethylene glycol (PEG) recovery in thefeces. Each participant was given 3 g polyethylene glycol (PEG) (E3350;Dow Chemical Co., Midland, Mich.) divided into 1 g doses at breakfast,lunch and dinner. Percent PEG recovery was measured in 24-h fecalcollections by turbidimetric assay and was used as a basis to excludesubject data in the case of poor compliance.

Gastrointestinal Symptoms

The presence of stomach noises, flatulence, bloating and abdominal painamong subjects was evaluated daily using a short questionnaire. Theseverity of gastrointestinal symptoms were assessed daily by self-reportusing a scale of 1-10 (0=none, 10=very severe) for 18 days during thesecond camp session.

Fractional Calcium Absorption Test

During the last week of each session, following an overnight fast, thesubjects participated in a calcium absorption test. On the morning ofthe test, phlebotomists inserted catheters and drew 10 ml baselinevenous samples. Immediately after the blood draw, participants consumeda breakfast consisting of an English muffin, scrambled eggs, butter andjam. The meal contained 150 mg calcium (Ca) from 2% milk plus 15 mg⁴⁴Ca, a stable non-radioactive isotope. The oral isotope wasadministered as a liquid (CaCl₂) which was added to the milk and allowedto equilibrate overnight. Following breakfast, participants were notallowed to consume any food but were allowed to drink ad libitumdeionized water. A second stable isotope, ⁴³Ca (3.5 mg) also as calciumchloride, was given intravenously one hour after the consumption ofbreakfast and the oral isotope. A final blood draw was performed threehours after administration of the intravenous dose after which catheterswere removed and subjects were served lunch.

Absorption and Retention Calculations

Two 24-h urine pools were used to measure changes in fractional calciumabsorption over 48 hours following calcium isotope administration. Urinesamples collected in 24-h pools for two days (0-24 h and 24-48 h) afterthe absorption test were analyzed for ⁴⁴Ca and ⁴³Ca enrichment by highresolution inductively coupled plasma mass spectrometry (ICP-MS,Finnegan Element2, Thermo Scientific). Calcium absorption (Equation 1)was calculated using enrichment values to calculate the A excess for⁴⁴Ca and ⁴³Ca as the difference of 0-24 h and 24-48 h samples frombaseline, divided by baseline. These excess values were then expressedas a ratio of ⁴⁴Ca/⁴³Ca based on their natural abundances and multipliedby the quantity of the iv dose (mg) divided by the oral dose (mg).

$\begin{matrix}{{Fractional}\mspace{14mu}{Ca}\mspace{14mu}{Absorption}{= \left( {\left\lbrack {\left( \frac{{\,^{44}{Ca}}\mspace{14mu}\Delta\mspace{14mu}{excess}}{{\,^{43}{Ca}}\mspace{14mu}\Delta\mspace{14mu}{excess}} \right)*\left( \frac{0.02083}{0.00135} \right)} \right\rbrack*\left\lbrack \frac{{iv}\mspace{14mu}{dose}}{{oral}\mspace{14mu}{dose}} \right\rbrack} \right)}} & {{Equation}\mspace{20mu} 1}\end{matrix}$

Balance data were used to calculate calcium retention (Equation 2) bysubtracting 24-h calcium excretion in urine and feces from 24-h dietarycalcium intake. The first 7 days of each 3-week study were regarded asan equilibration period for participants to acclimate to calcium intakeand the fiber treatment, while the remaining 2 weeks acted as theexperimental period. Balance was calculated based on as many of the 14days in the experimental portion as possible, so long as it allowedcalculations to begin and end on days with fecal samples. Periodsbetween stools were divided by the appropriate number of days. Dailyurinary calcium excretion values used in balance calculations werecorrected for variation in timing of collections and incompletecollections by adjusting 24 h urinary calcium for daily creatinineexcretion (Equation 3). Calcium retention was calculated using bothuncorrected and corrected values for urinary calcium excretion. Apparentcalcium absorption (Equation 4) was determined as the difference betweencalcium intake and fecal calcium excretion while net calcium absorptionefficiency (Equation 5) was calculated as intake minus fecal excretion,divided by intake.

$\begin{matrix}{{{Calcium}\mspace{14mu}{Retention}} = {{Dietary}\mspace{14mu}{Ca}\mspace{14mu}{Intake}\text{-}{Urinary}\mspace{14mu}{Ca}\text{-}{Fecal}\mspace{11mu}{Ca}}} & {{Equation}\mspace{14mu} 2} \\{{{Corrected}\mspace{14mu} 24h\mspace{14mu}{Urine}\mspace{14mu}{Ca}\mspace{14mu}{Excretion}} = \frac{24h\mspace{14mu}{Urinary}\mspace{14mu}{Ca}\mspace{14mu}({mg})}{\begin{matrix}\left\lbrack {24h\mspace{14mu}{Creatinine}\mspace{14mu}{({mg})/{Avg}}\mspace{14mu} 24h\mspace{14mu}{Creatine}} \right. \\\left. {{Excretion}\mspace{14mu}{for}\mspace{14mu}{Balance}\mspace{14mu}{Period}\mspace{14mu}({mg})} \right\rbrack\end{matrix}}} & {{Equation}\mspace{14mu} 3} \\{{{Apparent}\mspace{14mu}{Ca}\mspace{14mu}{Absorption}} = {{Dietary}\mspace{14mu}{Ca}\mspace{14mu}{Intake}\text{-}{Fecal}\mspace{14mu}{Ca}\mspace{14mu}{Excretion}}} & {{Equation}\mspace{14mu} 4} \\{{{Net}\mspace{14mu}{Ca}\mspace{14mu}{Absorption}\mspace{14mu}{Efficiency}} = {\frac{\begin{matrix}{{{Dietary}\mspace{14mu}{Ca}\mspace{14mu}{Intake}\mspace{14mu}({mg})} -} \\{{Fecal}\mspace{14mu}{Ca}\mspace{14mu}({mg})}\end{matrix}}{{Dietary}\mspace{14mu}{Ca}\mspace{14mu}{Intake}}*100}} & {{Equation}\mspace{14mu} 5}\end{matrix}$

Statistical Analysis

Statistical analyses were performed using SAS (version 9.2; SASInstitute, Cary, N.C.). Baseline characteristics of females and maleswere compared using t-tests. Wilcoxon's rank-sum test was used to assessdifferences in nonparametric gastrointestinal symptoms. Pearson'scorrelations were used to examine potential associations between thechange in fractional Ca absorption in 24-48 h urine (absorption on SCFminus absorption on control) and the differences in calcium balance andvitamin D status, baseline anthropometrics, and measures of bone densityand strength. A general linear model was used to assess the effect ofSCF on fractional calcium absorption. The model accounted for thecrossover design by nesting id within sequence as a random effectvariable and controlled for phase (first versus second 3-week campsession) and sequence of treatments. The data were analyzed separatelyfor each time period (0-24 h and 24-48 h). Similar analyses wereperformed for calcium balance. Using published means and standarddeviations for fractional calcium absorption reported in adolescentchildren, it was determined that a sample size of 24 would providesufficient power (80%) to see a 5.9% difference in calcium absorptionassuming an alpha error of 0.05 and a standard deviation of 2.9±9.6%.P-values <0.05 were considered statistically significant for allstatistical tests.

Results

A total of 24 subjects (9 girls and 15 boys) participated in this study.Three subjects did not participate in the fractional calcium absorptiontest in both sessions. Therefore, fractional calcium absorption wasanalyzed for 21 subjects. All other analyses included all available datavalues. The participants evaluated in this study were ethnically diversewith a distribution of 11 Asian, 6 Hispanic, 1 Black, and 6 multi-racialteens (Other). Subject characteristics including age, anthropometrics,physical characteristics, and bone measures are given in Table 1 whichpresents means and standard deviations for girls and boys separately.Girls and boys had similar physical characteristics; there werestatistically significant differences only in % lean (girls lower thanboys, P=0.009) and % fat mass (girls higher than boys, P=0.01) in thiscohort. Mean habitual intakes of calcium and dietary fiber were 768±403mg/d and 12±4 g/d, respectively.

TABLE 1 Baseline subject characteristics Females (n = 9) Males (n = 15)Age (y) 13.3 ± 1.0  13.5 ± 0.9  Weight (kg) 59.9 ± 13.2 61.1 ± 11.8Height (cm)* 157.3 ± 4.9   164.9 ± 8.2   BMI (kg/m²) 24.1 ± 4.0  22.4 ±3.1  BMI Percentile (%) 80.0 ± 16.4 74.7 ± 19.2 Tanner Score Average**3.8 ± 0.7 2.4 ± 0.9 Lean mass (%)** 61.2 ± 5.2  71.1 ± 9.5  Fat mass(%)* 35.1 ± 5.6  25.1 ± 10.0 Total Body BMD (g/cm²) 1.07 ± 0.11 1.04 ±0.11 Total Body BMC (g) 2115 ± 329  2316 ± 424  Total Spine BMD (g/cm²)1.09 ± 0.13 1.04 ± 0.14 Femoral Neck BMD (g/cm²) 1.03 ± 0.18 1.05 ± 0.15t-test; Mean ± SD *Significant difference between boys and girls (p <0.05) **Significant difference between boys and girls (p < 0.01)

Gastrointestinal Symptoms

No significant differences in the severity of gastrointestinal symptoms,including bloating, flatulence, abdominal cramping, and stomach noiseswere seen between SCF and Control treatments over the 18 day observationperiod (Table 2).

TABLE 2 Self-reported mean daily gastrointestinal symptoms over 18 daysin response to SCF consumption. Soluble Corn Fiber Control Bloating 0.1± 0.3 0.3 ± 0.8 Flatulence 0.6 ± 0.8 1.1 ± 1.7 Abdominal Cramping 0.3 ±0.5 0.4 ± 0.7 Stomach Noises 0.2 ± 0.7 0.3 ± 0.9 Wilcoxon's rank-sumtest; Mean ± SD N = 23 P > 0.05 for all parameters Scores were assignedbased on a 10 point scale: 0 = none, 1 = very mild, and 10 = verysevere.Fractional Calcium Absorption. Calcium Balance and Bone Biomarkers

Fractional calcium absorption was based on analysis of isotopes excretedin urine collected between 0-24 and 24-48 hours after isotopeadministration (FIG. 1). Compared with the control, mean fractionalcalcium absorption did not differ during the first 24 h but wassignificantly higher after treatment with SCF (0.595±0.142 vs.0.664±0.129, respectively; P=0.02) at 24-48 h. The difference in meanfractional calcium absorption of 0.069 represents an 11.6% increase inabsorption with SCF treatment. A general linear model (FIG. 2)identified significant effects of treatment at 24-48 h (P=0.02) but notat 0-24 h (P=0.09) on fractional calcium absorption. Treatment had nosignificant effect on net calcium absorption, net absorption efficiency,fecal calcium excretion or calcium retention (Table 3). Neither urinenor fecal calcium excretion were correlated with fractional calciumabsorption. Bone turnover marker concentrations are reported in Table 4.No differences in serum alkaline phosphatase, phosphorus, calcium,parathyroid hormone, leptin, insulin-like growth factor (IGF)-1,IGF-binding protein-3, sclerostin, and urine n-telopeptide cross links,calcium and phosphorus resulted from SCF consumption.

TABLE 3 Effect of soluble corn fiber treatment on calcium absorption andretention in 23 adolescent boys and girls SCF Control P-value StableIsotope Analysis Fractional Calcium Absorption, 0.522 ± 0.110 0.497 ±0.108 0.09  0-24 h Fractional Calcium Absorption, 0.664 ± 0.129 0.595 ±0.142 0.02 24-48 h Balance Calcium Intake (mg/d) 606 ± 29  604 ± 25 0.69 Urine Calcium (mg/d) 77 ± 56 65 ± 36 0.11 Fecal Calcium (mg/d) 318± 108 312 ± 106 0.77 Net Absorption Efficiency (%) 47 ± 18 48 ± 17 0.75Retention (mg/d) 212 ± 117 227 ± 101 0.42 General linear model thatincludes treatment, sequence, and phase Mean ± SD N = 21 for fractionalcalcium absorption values

TABLE 4 Serum and urine bone turnover marker concentrations at baselineand after treatment with 0 and 12 g SCF Bone Biomarker SCF ControlP-value Serum Alkaline Phosphatase, 235.57 ± 135.45  235.00 ± 132.5 0.92 U/L Calcium, ng/dl 10.23 ± 0.40  10.24 ± 0.40  0.72 Creatinine,ng/dl 0.85 ± 0.12 0.85 ± 0.12 0.88 Phosphorus, ng/dl 4.76 ± 0.59 4.76 ±0.58 0.21 Parathyroid Hormone, 21.23 ± 11.27 21.23 ± 11.02 0.82 pg/mlLeptin, ng/ml 8.65 ± 7.61 8.46 ± 7.50 0.89 IGF-1, 266.81 ± 52.30  269.59± 52.93  0.87 IGF-Binding Protein 3 3648.99 ± 572.71  3651.26 ± 560.23 0.46 Sclerostin, ng/ml 0.41 ± 0.17 0.41 ± 0.17 0.47 Urine N-telopeptide5378.81 ± 4632.47 5323.75 ± 4538.67 0.63 Crosslinks, nm BCE Calcium,mg/dl 3.43 ± 2.51 3.37 ± 2.47 0.69 Phosphorus, mg/dl 77.85 ± 39.97 76.69± 39.51 0.66 Creatinine, mg/dl 90.297 ± 45.721 90.67 ± 44.75 0.56 Mean ±SD

Vitamin D Status and Calcium Absorption

Mean vitamin D status after SCF and Control treatments was 65.2±18.8 nMand 59.1±15.9 nM, respectively, which were not significantly different.No significant associations between differences in 25-hydroxyvitamin Dand fractional calcium absorption (between 24-48 h) or net absorptionefficiency were observed.

Predictors of the Effect of SCF on Calcium Absorption

The difference in fractional calcium absorption between SCF and Controltreatments in 24-48 h urine collections was not correlated with height(r=0.112, P=0.63), body surface area (r=0.012, P=0.96), weight(r=−0.022, P=0.92), habitual dietary fiber (r=0.150, P=0.54) and calcium(r=0.012, P=0.96) intakes, Tanner stage (r=−0.131, P=0.57), or BMI(r=−0.074, P=0.75).

Example 2 Fecal Processing and DNA Extraction

Microbial community composition and structure in feces was determined insamples collected at the beginning and end of each session for eachsubject of Example 1. Frozen fecal samples were weighed, thawed at 4°C., then sterilized double distilled water (twice the weight of thefecal samples) was added and samples were homogenized in a stomacher.Fecal slurries were stored at −20° C. until DNA was extracted. DNA wasextracted from 50-100 mg of fecal material using the FastDNA® SPIN kitfor Soil (MP Biochemicals, Irvine, Calif.). DNA quality was checkedusing a 0.7% agarose gel and Nanodrop 1000 spectrophotometer (ThermoScientific, Wilmington, Del.) and then quantified using a Nanodrop 3300fluorospectrometer (Thermo Scientific).

Microbial Community Composition using Pyrosequencing

The phylogenetic diversity of bacterial communities was determined using16S rRNA gene sequences obtained using 454 FLX titanium chemistry andRoche Genome Sequencer (454 Life Sciences-Roche, Branford, Conn.) andprimers that amplify the V3-V5 region of the 16S rRNA gene. Multiplesamples were run and differentiated using 10-bp tagged forward primers.Initial PCR from fecal samples extracts was performed using highfidelity Phusion DNA Polymerase (NEB) and amplicons were gel purified(QIAEX II Gel Extraction Kit, Qiagen). At the Purdue Genomics facilitiespurified amplicons were quantified by fluorometry after staining usingthe PicoGreen DNA Assay Kit and by qPCR, and equimolar amounts were usedfor 454 FLX titanium chemistry sequencing.

Statistical Analysis

The reads from pyrosequencing analysis were first pre-processed usingsoftware to remove primer tags and to remove low quality sequences.Sequence quality was considered low if the length was <400 bp or ifthere were mismatches or ambiguities in the forward primer sequence.Sequences were analyzed using the QIIME pipeline that includes softwarefrom many sources that allows Operational Taxonomic Unit (OTU) andtaxonomic assignment as well as a number of different beta and alphadiversity measures. Chimerslayer was used to remove chimeric sequences.The OTU assignments were made using the uclust method and furthestneighbor clustering with a 97% sequence similarity threshold.Representative OTU sequences were obtained after sequence alignmentusing PyNast and the Greengenes core set. Taxonomic assignments weremade using the RDP classifier at 80% confidence. Rarefaction analysiswas used to obtain an estimation of sequence coverage of the community.Alpha biodiversity estimations (e.g., Shannon and Chao1 indices) werecalculated to compare subjects, with the caveat that PCR is being usedto target the 16S rRNA gene results might have been biased anddifferences in sequence copy per genome would influence relativenumbers. Community composition comparisons were made using “FastUniFrac” analysis of both OTU and phylogenetic datasets.

Pearson's correlation analysis was used to look for associations betweenthe difference in fractional calcium absorption between treatments(24-48 h) and the difference in the presence of bacteria genera aftereach treatment. Bacteria used in these correlations were for bacteriawith genera average >0.001 (=0.1%) or taxa proportions that weresignificantly different in end samples based on t-tests which includedthe following bacterial taxa: Bifidobacterium, Other Coriobacteriaceae,Bacteroides, Barnesiella, Butyricimonas, Parabacteroides, Prevotella,Alistipes, Other Rikenellaceae, Enterococcus, Lactobacillus, OtherLactobacillaceae, Streptococcus, Clostridium, Eubacterium,Mogibacterium, Blautia, Anaerostipes, Coprococcus, Dorea. OtherLachnospiraceae, Roseburia, Other Clostridiales, OtherPeptostreptococcaceae, Sporacetigenium, Acetivibrio, Butyricicoccus,Faecalibacterium, Oscillibacter, Other Ruminococcaceae, Ruminococcus,Subdoligranulum, Dialister, Other Clostridia, Catenibacterium,Coprobacillus, Other Erysipelotrichaceae, Turicibacter, OtherFirmicutes, Other Bacteria, Escherichia/Shigella, Pseudomonas,Actinomyces, Other Streptococcaceae, Anaerofustis, and Anaerococcus.

Results Changes in Bacterial Community Composition

A total of 1,793,821 sequences were obtained using 454-Titaniumpyrosequencing (Roche Applied Science, Branford, Conn., USA) with anaverage of 19498 sequences (±7126) per sample, and a range between 8211and 41212 sequences per sample. There were no significant differences inthe number of sequences obtained for each subject when compared byeither treatment (P>0.05) or by time of collection (baseline vs. endsamples) (P>0.05). There were ten phyla, Actinobacteria, Bacteroidetes,Firmicutes, Proteobacteria, Cyanobacteria, Fusobacteria, TM7,Verrucomicrobia, Spirochaetes and Synergistetes represented in themicrobial communities of the 23 subjects. But in all samples more than99% of the sequences were from four phyla; Firmicutes was the mostdominant phylum with an average of 89.4% followed by Bacteroidetes(5.1%), Actinobacteria (4.9%) and Proteobacteria (0.5%).

At the phylum level, regardless of the inclusion of SCF in the clinicaldiet, the average relative proportion of Bacteroidetes significantlyincreased and Firmicutes decreased by the end of each session.Communities from subjects on SCF versus Control treatments weresignificantly different at the family level (FIG. 3). There was a higherproportion of Porphyromonadaceae (P=0.02) and other Clostridiales(P=0.009) after the SCF diet and lower Peptostreptococcaceae (P=0.04).There was a significant difference in relative population proportions inthe family Corynebactariaceae (P=0.02) at the beginning of the twotreatments. At the lowest level of resolution for these sequences therewere nine genera and four “other” groups that had average proportionsthat differed significantly (P<0.1) after the SCF versus Controltreatments (Table 5). The significant increases after the SCF diet werein the genera Parabacteroides (P<0.003), other Clostridiales (P=0.04)and other Ruminococcaceae (P<0.03) while significant decreases wereobserved for Enterococcus (P <0.03), Anaerofustis (P<0.05), Coprococcus(P<0.03) and other Peptostreptococcaceae (P<0.002). There was also anincrease in Bifidobacterium, Alistipes, Anaerococcus, Catenibacteriumand other Clostridia but the increases were not significant. Similarly,decreases in Rothia, other Streptococcaceae, Clostridium,Sporacetilgenium, Turicibacter and other TM7 genera incertae sedisoccurred with SCF consumption but the difference in community proportionwith CON was not significant.

TABLE 5 Comparison of average (± SEM) proportion of bacterial taxa insubject fecal samples at end of soluble corn fiber supplemented versuscontrol diets Taxon* SCF CON P-value Phylum: Actinobacteria Class:Actinobacteria; Order: Actinomycetales; Family: Micrococcaceae Genus:Rothia 0.001 ± 0.002% 0.016 ± 0.061% 0.065 Class: Actinobacteria; Order:Bifidobacteriales; Family: Bifidobacteriaceae Genus: Bifidobacterium5.26 ± 1.18% 4.38 ± 1.03% 0.095 Phylum: Bacteroidetes Class:Bacteroidia; Order: Bacteroidales; Family: Porphyromonadaceae Genus:Parabacteroides 3.58 ± 1.18% 0.83 ± 0.32% 0.003 Class: Bacteroidia;Order: Bacteroidales; Family: Rikenellaceae Genus: Alistipes 1.77 ±0.62% 0.57 ± 0.19% 0.060 Class: Other Bacteroidetes  0.01 ± 0.005% 0.037± 0.145% 0.063 Phylum: Firmicutes Class: Bacilli; Order:Lactobacillales; Family: Enterococcaceae Genus: Enterococcus 0.012 ±0.037% 0.590 ± 2.100% 0.027 Class: Bacilli; Order: LactobacillalesFamily: Other Streptococcaceae 0.001 ± 0.003% 0.004 ± 0.008% 0.078Class: Clostridia; Order: Clostridiales; Family: Clostridiaceae Genus:Clostridium 1.012 ± 2.234% 1.960 ± 2.778% 0.077 Class: Clostridia;Order: Clostridiales; Family: Eubacteriaceae Genus: Anaerofustis 0.006 ±0.002% 0.012 ± 0.004% 0.048 Class: Clostridia; Order: Clostridiales;Family: Incertae Sedis XI Genus: Anaerococcus 0.017 ± 0.007% 0.003 ±0.002% 0.064 Class: Clostridia; Order: Clostridiales; Family:Lachnospiraceae Genus: Coprococcus 0.68 ± 0.13% 1.15 ± 0.25% 0.027Class: Clostridia Order: Other Clostridiales 14.64 ± 1.63%  9.61 ± 0.69%0.013 Class: Clostridia; Order: Clostridiales; Family: Other 0.42 ±0.10% 1.00 ± 0.18% 0.001 Peptostreptococcaceae Class: Clostridia; Order:Clostridiales; Family: Peptostreptococcaceae Genus: Sporacetigenium 5.46± 1.18% 9.38 ± 1.92% 0.083 Class: Clostridia; Order: ClostridialesFamily: Other Ruminococcaceae 4.07 ± 1.02% 1.93 ± 0.45% 0.030 Class:Other Clostridia 0.225 ± 0.413% 0.124 ± 0.266% 0.064 Class:Etysipelotrichi; Order: Erysipelotrichales; Family: ErysipelotrichaceaeGenus: Catenibacterium 2.297 ± 6.656% 0.438 ± 1.155% 0.094 Class:Elysipelotrichi; Order: Erysipelotrichales; Family: ErysipelotrichaceaeGenus: Turicibacter 0.308 ± 0.630% 0.855 ± 1.817% 0.083 Phylum: TM7Class: Other TM7 0.001 ± 0.003% 0.005 ± 0.012% 0.078genera_incertae_sedis *Only taxa in which proportions at the enddiffered significantly (paired t-test, p-values < 0.1) are listed.

Fast UniFrac with jackknife analysis showed no differences in communitystructure due to diet despite processing the data using differentclassification criteria (e.g., OTUs versus phylogeny). Principalcoordinate analysis of weighted Unifrac of OTUs showed some separationbetween samples at the beginning and end of clinical sessions but thetwo treatments did not separate.

Correlations Between Bacteria Genera and Fractional Calcium Absorption

Changes (SCF treatment minus control) in fractional Ca absorptionmeasured in 24-48 h urine were negatively correlated (decreases inbacterial genera as calcium absorption with SCF increased) withActinomyces, Pseudomonas from phylum Actinobacteria and otherErysipelotrichaceae from phylum Firmicutes. Conversely, change infractional Ca absorption was positively correlated (increases inbacterial genera as calcium absorption with SCF increased) withBacteroidetes member Bacteroides as well as Butyricicoccus,Oscillibacter, and Dialister from the phylum Firmicutes

TABLE 6 Correlations between calcium absorption and bacteria genera thatmay affect lower gut mechanisms Gcnera differences at Diff in Ca Abs at24-48 h end of each treatment Coefficient P-value Bacteroides   0.4830.027 Actinomyces −0.553 0.009 Pseudomonas −0.473 0.03  Butyricicoccus  0.454 0.039 Oscillibacter   0.565 0.008 Dialister   0.619 0.003 OtherErysipelotrichaceae −0.463 0.034 Pearson's correlations N = 21

The above results indicate that daily consumption of 12 g soluble cornfiber for 21 days in adolescent girls and boys increased fractionalcalcium absorption by ˜12%. This increase in fractional calciumabsorption occurred between 24 and 48 h as the effect was significant bymeasuring the second 24-h urine pool (24-48 h) after receiving stablecalcium isotopes, while no significant difference was seen in isotopeenrichment in urine collected during the first 24-h. This is supportedby literature that suggests microbial involvement and lower gutabsorption is not captured until 24 h after receiving isotopes.

It is difficult to say whether the increased absorption of calcium seenin this study resulted in bone mineral deposition because no effect wasseen on calcium retention. Fecal calcium measures are highly variable; asample size of 34 would be necessary to see a 61 mg difference incalcium retained with an alpha error of 0.05, 80% power and a standarddeviation of the difference between participants of 122 mg/d. It ispossible that the treatment elicits an effect on bone strength. The twomethods have a large difference in % SD, i.e. 9.7% for breaking forceand 41.3% for calcium retention. Assuming that the increased absorptionof calcium (measured by a more sensitive dual isotope method) with SCFis retained, data from this study suggest that treatment with SCF wouldlead to additional calcium retention of 70 mg/d. Over a year this wouldaccount for an additional 25 g of calcium or 2.8% of total body calciumassuming that an adult skeleton has 900 g of calcium.

In conclusion, consumption of 12 g/d SCF positively influenced calciumabsorption in adolescent girls and boys. SCF-induced absorption occurredafter 24 hours which may be indicative of lower gut involvement.Significant increases were seen in proportions of members of theBacteroidetes and Bifidobacteria, fermenters of resistant starches.

Example 3 Subjects and Methods Methods

Gut microbiota composition in representative samples with (10 g/day dose(“D10”) and 20 g/day dose (“D20”)) and without (0 g/day dose (“DO”)) SCFdietary treatment were determined using Illumina MiSeq high throughputsequencing instead of 454 pyrosequencing. The data is used to determineproportional increases or decreases in populations that are associatedwith differences in diet supplementation. There are five steps for gutmicrobial community analysis that were performed: (1) Fecal samplescollected at the beginning and end of each randomly assigned diet SCFsupplementation were homogenized in preparation for DNA extractions (atotal of six samples per subject with the exception of subject 103 whodid not submit a DO beginning fecal sample); (2) Total fecal DNA wasextracted using the Fast DNA™ Soil Spin kit and FastPrep™ system; (3)The extracted DNA was subjected to PCR using primers that target theBacteria 16S rRNA gene; (4) The PCR products were sequenced using theIllumina MiSeq; (5) Sequences were analyzed using the QIIME pipeline todetermine quantitative changes in microbial community members resultingfrom soluble corn fiber treatment.

Fecal Processing and DNA Extraction

Frozen fecal samples were processed and the DNA was extracted asprovided in Example 2.

Microbial Community Composition

The phylogenetic diversity of bacterial communities was determined using16S rRNA gene sequences obtained from high throughput paired end MiSeqtechnology (Illumina), and primers that amplify the V3-V4 region of the16S rRNA gene were used. Multiple samples were run and differentiatedusing a combination of 8-bp tagged forward primer and 8-bp taggedreverse primers using a step out protocol that uses two PCR runs. Thefirst PCR specifically amplifies the 16S rRNA gene from fecal samplesextracts. Unincorporated primers and nucleotides were separated from PCRamplicons using Agencourt AMPure XP kit (Becker). The second PCR wasused to add bitags to the amplicons (from the first run) that are neededfor Illumina sequencing and purified again using the Agencourt AMPure XPkit. All PCR was performed using Q5® High Fidelity DNA Polymerase (NewEngland Biolabs) to minimize error rate during polymerization. Purifiedamplicons were quantified by fluorometry after staining using thePicoGreen DNA Assay Kit. Amplicons from each sample were combined inequivalent quantities sequenced using the MiSeq instrument (Illumina).

Sequence Analysis

Sequences were pre-processed to remove primer tags and low qualitysequences, and then analyzed using the QIIME pipeline. MiSeq Illuminasequences of 16S rRNA gene fragments were analyzed using both OTU-basedand taxonomy based on phylogenetic trees. OTU was defined as a groupbased strictly on sequence similarity and not aligned to a knowntaxonomy. Sequences were first prefiltered and OTU assignments madeusing the uclust method and the Greengenes core sequences using a 60%threshold value (as recommended by QIIME developers). Representative OTUsequences were obtained after sequence alignment using PyNast to filterout sequences that did not align with the Greengenes core sequences.Taxonomic assignments were made using the RDP classifier at 80%confidence and the Greengenes database. Rarefaction analysis was used toobtain an estimation of sequence coverage of the community. Alphabiodiversity estimations were calculated to compare microbiota diversitywithin subjects under specific SCF treatments. Beta diversitycomparisons between communities were made using “Fast UniFrac” analysisof phylogenetic distances as well as non-phylogenetic distance analysisusing Euclidean distances. All alpha and beta diversity measures weremade using equivalent number of taxa (based lowest number of sequencesobtained from a single sample) that were randomly chosen using multiplerarefaction results (10 iterations).

Statistical Analyses

Friedman analysis (non-parametric equivalent to ANOVA) was used for anoverall comparison of average proportions of genera in subjects atbeginning (B) and end (E) of each SCF treatment. The Wilcoxon signedranks test was then used to determine significant differences pairwisecomparisons of samples from the beginning and end of each treatmentphase as well as between end samples. Student's T-test was used todetermine significant differences between alpha diversity measures.Significant differences in beta diversity between communities weredetermined using perMANOVA a non-parametric multivariate statisticaltool available in the Paleontological Statistics package version 2.16(PAST software, http://folk.uio.no/ohammer/past/index.html). Bonferronicorrection was applied to all statistical tests.

Results

The number of fecal samples analyzed for the 28 individuals was 167, sixper individual with the exception of one subject who did not provide abeginning sample for the 0 g-diet supplementation experiment (Table 7).For this reason the statistical results presented in this report arebased on data from only 27 subjects. The subjects were administered 10g/day of SCF (D10), 20 g/day of SCF (D20), and no SCF (DO).

Number of Sequences

A total of 12,979,388 high quality merged sequences were obtained usingMiSeq Illumina sequencing with an average of 77,720.9 sequences(±28,401) per sample, and ranged from 28,854 to 262.312 sequences persample (Table 7). The lowest number of sequences obtained was 28,854therefore all subsequent analyses were rarefied to 28,800 sequences persample. To obtain a rarefied dataset, 10 iterations of randomly choosing28,800 sequences from each dataset was performed then datasets weremerge to obtain a set of 28,8000 sequences that were representative ofeach sample.

TABLE 7 Subjects included in the microbial community analysis and numberof sequences from each fecal sample collected. Subject ID B-D10* E-D10B-D20 E-D20 B-D0 E-D0 Mean 101 35648 95561 28854 81110 39956 12786868166 102 32756 88022 53025 79399 39773 102533 65918  103** 46731 8363962571 57150 ND 114458 72910 105 71802 87419 77341 97803 78244 13506391279 106 47326 78151 33267 92006 43006 98303 65343 107 41246 8431534342 77309 37403 107778 63732 108 69813 71916 104705 58411 76103 7072175278 109 48771 90250 32212 59657 39759 64715 55894 110 104086 8054070670 87591 81174 102184 87708 111 40890 74227 39385 86600 39783 8415960841 112 37436 83953 38347 65681 42727 79281 57904 113 44561 7328741348 67915 30805 53853 51962 114 82124 100597 70023 99104 67147 9332385386 115 42685 85418 48484 85940 54404 93652 68431 116 55328 10419862443 99053 38824 99359 76534 117 45551 70450 66727 119448 40043 7832570091 118 70595 86264 85866 96618 83459 98271 86846 119 46358 8046647594 106314 46887 98310 70988 120 57301 96805 66532 101655 52993 12409083229 121 53277 72966 53345 76849 66119 137385 76657 122 76244 10577788763 119395 71736 81078 90499 124 86440 112472 73716 94871 112156 7960793210 125 92836 90916 83968 81214 69313 98037 86047 126 101758 12771487905 89984 110822 101913 103349 127 108740 88143 85892 94263 10397059492 90083 128 93626 95191 56287 77558 68134 89239 80006 129 8103493014 81428 75555 56459 48613 72684 130 262312 66058 87767 116323 95907118085 124409 *B- denotes the beginning sample, and E- denotes the endsample. **excluded from statistical analyses because of missing sample

Comparison of Phyla Represented in Sequence Data

There were 13 phyla, Actinobacteria, Bacteroidetes, Firmicutes,Proteobacteria, Chloroflexi, Cyanobacteria, Fusobacteria, Lentisphaerae,Synergistetes, TM7, Tenericutes, [Thermi] and Verrucomicrobia found inthe microbial communities of the 28 subjects. In addition, the primersamplified some sequences from the domain Archaea and other Bacteria thatcurrently cannot be classified. But more than 99% of the sequences werefrom four phyla, Actinobacteria, Bacteroidetes, Firmicutes andProteobacteria. Across all samples from all subjects the Firmicutes wasthe dominant phylum with an average of 65.8% followed by Bacteroidetes(26.0%), Actinobacteria (6.2%) and Proteobacteria (1.8%) (FIG. 1). Therewas no significant difference in proportions of phyla between thebeginning and end samples from any of the SCF test treatments. Therewere also no significant differences found at the Class and Order levelof taxonomic classification. Archaea are an important group that shouldbe monitored in the future but because the primers used in this studywere not developed for their inclusion we do not think it is proper touse their presence (or absence) in our assessment.

Comparison of Families Represented in Sequence Data

ANOVA of the taxa at the family level indicated there was a significantdifference only within the phylum Bacteroidetes. The Bacteroidetesincluded the families Bacteroidaceae, Porphyromonadaceae,Prevotellaceae, and Rikenellaceae and tentative new families[Barnesiellaceae], [Odorlbacteraceae], [Paraprevotellaceae],[Weeksellaceae], RF16, S24-7, and three other families. Families listedas [tentative] and “other” are those have yet to be officiallyclassified, mainly because these groups are recent discoveries based onmolecular analyses of which some have no representatives in culture touse for taxonomic assignment. ANOVA indicated that there weresignificant differences in the Porphyromonadaceae that was supported byBonferroni correction (p<0.0001).

Comparison of Genera Represented in Sequence Data

For further resolution the same comparisons were made at the genus levelof phylogenetic classification using non-parametric statistics. Of the235 genera (or genera equivalents) identified in analysis of all subjectsamples that were sequenced, a subset of only 24 genera comprised >1% ofthe community in at least one of the samples and represented about 90%of the communities (results not shown). Although some genera made up asmall proportion of the communities they did differ significantly.Genera that differed significantly were Parabacteroides, Bacteroides,Dorea, Lachnospira, an unclassified Ruminococcus, unclassifiedLachnospiraceae and “other” Bacteria (Table 8) based on Friedmananalysis (non-parametric equivalent to ANOVA) with Bonferonnicorrection.

TABLE 8 Friedman analysis (non-parametric equivalent to ANOVA) ofaverage proportions of genera* in subjects at beginning (B) and end (E)of each SCF treatment (10, 20, and 0 g/day) B-D10 E-D10 B-D20 E-D20 B-D0E-D0 Bonferroni (%) (%) (%) (%) (%) (%) P Corrected P Parabacteroides0.90 2.11 1.12 3.01 0.99 1.06 0.0000 0.0000 Uncl. 6.35 11.39 6.04 13.085.87 6.32 0.0000 0.0000 Lachnospiraceae 3.75 2.38 2.48 1.89 3.58 3.740.0000 0.0000 Reclassified [Ruminococcus] 1.08 1.04 1.22 0.70 1.14 1.260.0000 0.0015 Dorea Other_Bacteria 0.07 0.05 0.12 0.06 0.11 0.04 0.00000.0016 Lachnospira 1.14 0.67 1.43 0.66 1.26 0.94 0.0000 0.0067Bacteroides 14.75 11.09 16.18 9.62 15.64 13.74 0.0001 0.0216Turicibacter 0.22 0.12 0.12 0.13 0.15 0.27 0.0011 0.2545 Dialister 0.640.86 0.74 1.17 0.55 0.46 0.0016 0.3796 SMB53 2.46 1.92 1.89 1.38 2.081.93 0.0026 0.6035 Ruminococcus 5.12 7.15 5.89 7.70 5.00 5.94 0.00410.9555 Streptococcus 0.39 0.18 0.19 0.46 0.37 0.22 0.0048 1Adlercreutzia 0.06 0.05 0.06 0.04 0.06 0.06 0.0057 1 Butyricimonas 0.130.10 0.17 0.16 0.17 0.18 0.0089 1 Porphyromonas 0.00 0.01 0.00 0.00 0.000.01 0.0094 1 Other_Clostridiaceae 0.32 0.09 0.30 0.11 0.25 0.26 0.01511 Fusobacterium 0.000 0.001 0.001 0.000 0.000 0.001 0.0156 1 UnclAlcaligenaceae 0.001 0.002 0.001 0.002 0.001 0.001 0.0162 1 Prevotella5.73 6.35 6.30 4.72 6.70 6.64 0.0168 1 Varibaculum 0.000 0.001 0.0020.001 0.000 0.001 0.0170 1 Uncl. Erysipelotrichaceae 0.01 0.01 0.01 0.000.01 0.01 0.0231 1 Akkermansia 0.000 0.003 0.001 0.001 0.001 0.0010.0232 1 Campylobacter 0.001 0.001 0.002 0.001 0.000 0.001 0.0233 1Granulicatella 0.003 0.001 0.001 0.001 0.003 0.002 0.0241 1 Sutterella0.80 0.79 1.01 0.93 1.10 1.06 0.0245 1 Uncl. Enterobacteriaceae 0.220.36 0.26 0.15 0.24 0.47 0.0255 1 Corynebacterium 0.00 0.00 0.00 0.010.06 0.00 0.0297 1 Acidaminococcus 0.10 0.12 0.05 0.15 0.07 0.10 0.03011 Odoribacter 0.39 0.29 0.45 0.88 0.36 0.50 0.0304 1 WAL_1855D 0.00 0.000.00 0.00 0.00 0.00 0.0312 1 Anaerostipes 0.22 0.14 0.18 0.13 0.33 0.220.0344 1 Abiotrophia 0.00 0.00 0.00 0.00 0.00 0.00 0.0423 1 *Only generathat differed significantly (p < 0.05) are included in the table “Uncl(Unclassified)” designation indicates that the sequence likely belongsto a new genus that has yet to be described but there is sufficientinformation that supports the identification of a new genus. “Other”designation also for unclassified taxa but at higher taxonomic levels,i.e., class, order or family.

Pairwise comparison (Wilcoxon Signed Rank test with Bonferronicorrection) of proportional averages of all genera in the beginning andend samples collected within SCF treatments were made to identify thetreatments in which these taxa differed significantly. Proportions ofParabacteroides and an unclassified Lachnospiraceae (Table 8) weresignificantly greater at the ends of diets D10 and D20 compared to thebeginning. In addition, at the end of diet D10 there was a significantincrease in Akkermansia and decrease in reclassified [Ruminococcus]. Atthe end of diet D20 there were significant decreases in Bacteroides andLachnospira. At the end of diet DO there was a significant decrease in“other” Bacteria but this is not a single taxonomic group. Therefore,unlike the other two diet treatments there were no significantdifferences in taxa in subjects after consuming diet DO.

TABLE 8 Comparison of average proportions (%) of genera in subjects thatsignificantly differed (Wilcoxon signed rank test after Bonferronicorrection, p < 0.05) between the beginning (B) and end (E) of each SCFtreatment (10, 20, and 0 g/day) Phylum/Genus B-D10 E-D10 p B-D20 E-D20 pB-D0 E-D0 p Bacteroidetes Bacteroides 14.7 11.1 ns 16.2 9.6 0.011 15.613.7 ns Parabacteroides 0.9 2.1 0.002 1.1 3.0 0.001 1.0 1.1 nsFirmicutes Lachnospira 1.1 0.7 ns 1.4 0.7 0.002 1.3 0.9 ns UnclassifiedLachnospiraceae 6.4 11.4 0.001 6.0 13.1 0.0003 5.9 6.3 ns reclassifiedRuminococcus 3.7 2.4 0.011 2.5 1.9 ns 3.6 3.7 ns VerrucomicrobiaAkkermansia 0.000 0.003 0.020 0.001 0.001 ns 0.001 0.001 ns OtherBacteria 0.07 0.05 ns 0.12 0.06 ns 0.11 0.04 0.001

Furthermore, pairwise comparisons of diets D10, D20, and DO end samplessubstantiated the differences in communities (Table 9). There was apotential dosage effect on the Parabacteroides, this was indicated bysignificantly greater proportions after diet D20 compared to D10 thatwas greater than after diet DO. The same trend was found for theunclassified Lachnospiraceae and Dialister, which were significantlygreater at the end of diets D10 and D20 compared to DO but diets D10 andD20 were not significantly different. Also, there was a significantlygreater proportion of Bifidobacterium at the end of diet D20 compared toDO. At the end of diets D10 and/or D20 compared to DO there weresignificantly lower proportions of Anaerostipes, Dorea, reclassified[Ruminococcus], and unclassified Erysipelotrichaceae. At the beginningof SCF diet treatment there were also significant differences inproportions of reclassified [Ruminococcus], Enterococcus andCampylobacter. The significantly greater proportion of reclassified[Ruminococcus] at the beginning of diet D20 potentially factored intofinding a significant difference in this taxon in the comparison of thebeginning and end samples.

TABLE 9 a: Comparison of average proportions of genera* that differedsignificantly at the beginning (B) of each SCF treatment (10, 20, and 0g/day) using Bonferroni corrected Wilcoxon Sum Rank Test(non-parametric) B-D10 B-D20 B-D0 Phylum Genus (%) (%) (%) FirmicutesReclassified 3.75  2.48  3.58  Ruminococcus Firmicutes Enterococcus0.001 0.027 0.009 Proteobacteria Campylobacter 0.001 0.002 0.000 b:Comparison of average proportions of genera* in subjects at end (E) ofeach SCF treatment (10, 20, and 0 g/day) using Bonferroni correctedWilcoxon Sum Rank Test (non-parametric) E-D10 E-D20 E-D0 Phylum Genus(%) (%) (%) Actinobacteria Bifidobacterium  4.23   5.06  3.24 Bacteroidetes Parabacteroides  2.11   3.01  1.06  FirmicutesAnaerostipes  0.13   0.13  0.22  Firmicutes Dorea  1.04   0.70  1.26 Firmicutes Reclassified  2.38   1.89  3.74  Ruminococcus FirmicutesUnclassified 11.39  13.08  6.32  Lachnospiraceae Firmicutes Dialister 0.86   1.17  0.46  Firmicutes Unclassified  0.006  0.004 0.008Erysipelotrichaceae

Without being bound to a particular theory, the increase in proportionof Parabacteroides, unclassified Lachnospiraceae and Dialister afterdiets D10 and D20 suggests that these microbes are involved in SCFfermentation.

Comparison of Alpha Diversity

There were significant differences (p<0.05) in alpha diversity measuresbetween the beginning and end communities under each SCF diet (Table10). The alpha diversity provides a metric for diversity within atreatment. Using the Chao1 measure these differences were found for bothSCF diets D10 and D20 and not DO (Table 10, FIG. 5). Whereas usingObserved Species the difference was only significant for diet D20. Nosignificant differences were found for PD Whole Tree (Table 10a).Whereas pairwise comparison of the diversity in the end samplesindicated there were significant differences between all Chao1 valuesand between end of D10 and D20 versus DO. The differences insignificance among the diversity indices tested are because thealgorithms for each of these alpha diversity measures are quitedifferent with emphasis on different criteria. For example PD whole treeis a phylogenetic measures whereas the other two are not. Chao1 is ameasure of species richness and observed species sums up the number ofunique OTUs. Without being bound to a particular theory, it is believedthat the SCF diet was increasing the number of taxa in the samples.

TABLE 10a Comparison of mean ± standard deviation of Alpha diversityvalues. Significant differences between beginning and end within eachtreatment B-D10 E-D10 B-D20 E-D20 B-D0 E-D0 Chao 1 1141 ± 155 1282 ± 1921146 ± 154 1402 ± 276 1152 ± 186 1104 ± 126 PD whole tree 30.8 ± 4.231.2 ± 4.2 31.2 ± 4.1 31.7 ± 3.7 30.8 ± 4.2 29.8 ± 4.4 Observed Species624.8 ± 82.3 634.5 ± 83.8 619.6 ± 91.4 649.6 ± 75.5 615.1 ± 83.4 601.4 ±83.5

TABLE 10b Significant differences between beginning and end sampleswithin each treatment B-D10 E-D10 B-D20 E-D20 B-D0 E-D0 Chao 1 1141 ±155 1146 ± 154 1152 ± 186 1282 ± 192 1402 ± 276  1104 ± 126^(c) PD wholetree 30.8 ± 4.2 31.2 ± 4.1 30.8 ± 4.2 31.2 ± 4.2 31.7 ± 3.7 29.8 ± 4.4Observed Species 624.8 ± 82.3 619.6 ± 91.4 615.1 ± 83.4 634.5 ± 83.8649.6 ± 75.5 601.4 ± 83.5

Community Comparisons Using Beta Diversity

Comparisons between communities (beta diversity) revealed separation ofsome communities dependent on the distance measures used.Non-phylogenetic Euclidean distances (Binary Euclidean and Bray Curtis)and phylogenetic distances (Unifrac G, Unifrac weighted, and Unifracunweighted) with jackknife were tested to determine differences incommunity structure between samples and factors likely contributing tothese differences. Principal Coordinate Analysis (PCoA) clustering ofnon-phylogenetic Euclidean distances indicated communities at the end ofSCF treatments D10 and D20 separated from those at the end of the DO andall the beginning samples (FIGS. 6 and 7). Separation was most evidentusing binary Euclidean distances (FIG. 7). Separation can be seen acrossthe first PCoA axis that accounted for 12.11% of variation and to someextent along the second PCoA axis that accounts for 9.48% of variation.Whereas using any of the Unifrac phylogenetic distances clustering ofsamples were more by subject rather than by treatment (example UnifracG, FIG. 8). This indicates that the phylogenetic composition ofcommunities is more similar within a subject than between subjects. Thisis similar to previous reports of high variation between the gutmicrobiome of humans. These Euclidean and phylogenetic distances arecalculated using different criteria that provide insight into factorscontributing to differences in gut microbial communities. For example,Euclidean distances are based on presence or absence of every OTU(operational taxonomic unit) in each community. This indicates that thepresence or absence of specific taxa is contributing to the differencesin the community.

Non-Parametric Permutation Multivariate ANOVA

Non-parametric permutation multivariate ANOVA (perMANOVA) afterBonferroni correction revealed significant differences betweentreatments that were seen as clusters in the Principal Coordinate (PCoA)scatterplots of beta diversity analysis. Significant differences werefound between communities in samples from the beginning and end samplesof diets D10 and D20 with their respective beginning samples using theEuclidean distances measured (Bray Curtis, binary Euclidean) but not thephylogenetic measures (Unifrac distances) (Table 11, results also shownin FIGS. 6-8). There were also significant differences in the Euclideanand Bray Curtis distances between the end samples of diet D20 comparedto the end DO. End samples of diets D10 and DO also differedsignificantly but only using Euclidean distances. There were alsodifferences in the Unifrac G distances but because differences were alsofound between beginning samples it may not be a result of the SCFtreatments. Prior to Bonferroni corrections there were more significantdifferences (Table 11) but here we have chosen to focus on the morestringent cutoff. However, the data has been included in the reportbecause the stringency of Bonferroni can include false negatives.Regardless, these results clearly illustrates that the microbialcommunities of the subjects at the end of SCF diet treatment D20differed the most, suggesting that it was the code for the highest SCFdose given to the subjects.

TABLE 11 Summary of per MANOVA using various beta distance measures ofcommunities (average proportion of genera) between the beginning (B) andend (E) of each SCF treatment (10, 20, and 0 g/day) B-D10 B-D20 B-D10E-D10 Distance v E- v E- B-D0 v v B- B-D10 B-D20 v E- E-D10 E-D20Measure D10 D20 E-D0 D20 v B-D0 v B-D0 D20 v E-D0 v E-D0 Bray-Curtis0.0003 0.0000 1.0000 0.9999 1.0000 0.9999 0.9994 0.0038 0.0000 Euclidean0.0000 0.0000 0.9999 0.9998 1.0000 0.9985 0.9778 0.0004 0.0000 Unifrac g0.3002 0.2936 0.0056 0.0106 0.0115 0.0020 0.0311 0.0000 0.0000 Unifrac0.4507 0.0360 0.8434 1.0000 1.0000 0.9984 0.9999 0.9538 0.3064unweighted Unifrac 0.3544 0.0133 0.9876 0.9030 0.9863 0.9878 0.82030.3676 0.0468 weighted With Bonferroni correction Bray-Curtis 0.00510.0002 1.0000 1.0000 1.0000 1.0000 1.0000 0.0575 0.0003 Euclidean 0.00030.0002 1.0000 1.0000 1.0000 1.0000 1.0000 0.0054 0.0002 Unifrac g 1.00001.0000 0.0846 0.1590 0.1727 0.0303 0.4670 0.0003 0.0002 Unifrac 1.00000.5400 1.0000 1.0000 1.0000 1.0000 1.0000 1.0000 1.0000 unweightedUnifrac 1.0000 0.1992 1.0000 1.0000 1.0000 1.0000 1.0000 1.0000 0.7026weighted

The foregoing description of embodiments of the present invention hasbeen presented for the purposes of illustration and description. It isnot intended to be exhaustive or to limit the invention to the preciseforms disclosed. As the person of skill in the art will recognize, manymodifications and variations are possible in light of the aboveteaching. It will be apparent to those skilled in the art that variousmodifications and variations can be made to the present inventionwithout departing from the scope of the invention. Thus, it is intendedthat the present invention cover the modifications and variations ofthis invention provided they come within the scope of the claims andtheir equivalents.

1-129. (canceled)
 130. An edible composition comprising soluble cornfiber and one or more bacterial populations selected from the generaButyricicoccus and Oscillibacter, wherein the soluble corn fiber is madeby a process comprising: providing an aqueous feed composition that is adextrose syrup or a syrup made by hydrolysis of corn starch, the aqueousfeed composition having a solids concentration of at least 90% byweight; and contacting the feed composition for a time in the range of0.1-15 minutes at a temperature of at least 149° C. with at least onecatalyst that accelerates the rate of cleavage or formation of glucosylbonds, wherein the at least one catalyst is an acid added to the feedcomposition in an amount sufficient to make the pH of the feedcomposition 1.0-2.5, to form a product composition that is at least 50%by weight on a dry solids basis slowly digestible as identified as beingslowly digestible or digestion resistant by the Englyst Assay, theproduct composition including less than 50% on a dry solids basis ofresidual monosaccharides.
 131. The edible composition according to claim130, wherein the one or more bacterial populations are selected from thegenus Butyricicoccus.
 132. The edible composition according to claim130, wherein the one or more bacterial populations are selected from thegenus Oscillibacter.
 133. The edible composition according to claim 130,comprising two or more bacteria populations, each from a different genusselected from Butyricicoccus and Oscillibacter.
 134. The ediblecomposition according to claim 1, wherein the composition comprises atleast 2.5 g of soluble corn fiber per serving.
 135. The ediblecomposition according to claim 1, wherein the composition comprises nomore than about 100 g of soluble corn fiber per serving.
 136. A methodfor increasing one or more colonic bacteria populations in a mammaliansubject selected from the genera Butyricicoccus and Oscillibactercomprising administering an edible composition according to claim 1.137. The edible composition for use according to claim 136, wherein thesoluble corn fiber is administered at a rate of at least about 10 g/day,over the course of at least two weeks.
 138. A method for increasing oneor more colonic bacteria populations in a mammalian subject selectedfrom the genera Butyricicoccus and Oscillibacter comprisingadministering to the mammalian subject a composition comprising solublecorn fiber.
 139. The method according to claim 138, wherein the solublecorn fiber is made by process comprising: providing an aqueous feedcomposition that is a dextrose syrup or a syrup made by hydrolysis ofcorn starch, the aqueous feed composition having a solids concentrationof at least 90% by weight; and contacting the feed composition for atime in the range of 0.1-15 minutes at a temperature of at least 149° C.with at least one catalyst that accelerates the rate of cleavage orformation of glucosyl bonds, wherein the at least one catalyst is anacid added to the feed composition in an amount sufficient to make thepH of the feed composition 1.0-2.5, to form a product composition thatis at least 50% by weight on a dry solids basis slowly digestible asidentified as being slowly digestible or digestion resistant by theEnglyst Assay, the product composition including less than 50% on a drysolids basis of residual monosaccharides.
 140. The method according toclaim 139, wherein at least one of the one or more increased bacterialpopulations is from the genus Butyricicoccus.
 141. The method accordingto claim 139, wherein at least one of the one or more increasedbacterial populations is from the genus Oscillibacter.
 142. The methodaccording to claim 139, wherein the soluble corn fiber is administeredat a rate of at least about 10 g/day, over the course of at least twoweeks.
 143. The method according to claim 140, wherein the soluble cornfiber is administered at a rate of at least about 10 g/day, over thecourse of at least two weeks.
 144. The method according to claim 141,wherein the soluble corn fiber is administered at a rate of at leastabout 10 g/day, over the course of at least two weeks.